Method and composition for detection and treatment of breast cancer

ABSTRACT

The present invention provides a method for the detection of breast cancer using breast by measuring expression levels of breast cancer specific marker (BCSM) genes, and in particular the level of polynucleotides transcribed from and polypeptides encoded by the BCSM genes. The present invention also provide a method for the treatment and/or prevention of breast cancer by modulating the activity of BCSM genes or the products of BCSM genes.

RELATED APPLICATION

[0001] This application is related to U.S. Provisional ApplicationSerial No. 60/359,999, filed Feb. 28, 2002.

TECHNICAL FIELD

[0002] The present invention relates generally to the detection andtreatment of cancer, and in particular breast cancer. The inventionspecifically relates to breast cancer-specific genes (BCSG), and topolynucleotides transcribed from and polypeptides encoded by the BCSGs.Such polynucleotides and polypeptides may be used for the detection andtreatment of breast cancer.

BACKGROUND

[0003] Breast cancer is the second leading cause of cancer-relateddeaths of women in North America. Although advances have been made indetection and treatment of the disease, breast cancer remains the secondleading cause of cancer-related deaths in women, affecting more than180,000 women in the United States each year.

[0004] Approximately 10% of all breast cancers are currently classifiedas strongly familial with many of these appearing to be caused bymutations in the hereditary breast cancer genes BRCA1 or BRCA2. However,at least one-third of breast cancers that seem to run in families arenot linked to BRCA1 or BRCA2, suggesting the existence of an additionalhereditary breast cancer gene or genes. Recently, structural andfunctional studies of cancer cell lines and tissues have demonstratedthe involvement of many genetic loci and genes in the development ofhuman breast cancer. Cytogenesis and loss of heterozygocity (LOH)studies have led to the discoveries of alterations in human chromosomesincluding 1p, 1q, 3p, 6q, 7q, 11p, 13q, 16q, 17p, 17q, and 18q, atfrequencies as high as 20-60%. Thus, multiple genes are involved in thedevelopment of extensively heterogeneous breast cancers.

[0005] No vaccine or other universally successful method for theprevention or treatment of breast cancer is currently available.Management of the disease currently relies on a combination of earlydiagnosis (through routine breast screening procedures) and aggressivetreatment, which may include one or more of a variety of treatments suchas surgery, radiotherapy, chemotherapy and hormone therapy. The courseof treatment for a particular breast cancer is often selected based on avariety of prognostic parameters, including an analysis of specifictumor markers. (See, e.g., Porter-Jordan and Lippman, Breast Cancer8:73-100, 1994). However, the use of established markers often leads toa result that is difficult to interpret, and the high mortality observedin breast cancer patients indicates that improvements are needed in thetreatment, diagnosis and prevention of the disease.

[0006] Accordingly, there is a need in the art for improved methods fortherapy and diagnosis of breast cancer. The identification of expressionprofiles and differentially expressed genes in the genomic scale wouldgreatly facilitates the molecular classification of tumors and discoveryof genes that are causally related to breast cancer development.

SUMMARY OF THE INVENTION

[0007] The present invention provides compositions and methods for thediagnosis and treatment of breast cancer. Specifically, the presentinvention discloses genes that are differentially expressed in breastcancer cell lines and breast cancer tissue samples as compared tocontrol cell lines and normal tissue samples, the polynucleotidestranscribed from these genes (SEQ ID NOS:1-19), and the polypeptidesencoded by these polynucleotides (SEQ ID NOS:20-38). The differentiallyexpressed genes are designated as breast cancer specific genes (BCSG).The polynucleotides transcribed from and the polypeptides encoded by theBCSGs are designated as breast cancer specific markers (BCSM).

[0008] In one aspect, the present invention provides a method fordiagnosing and monitoring breast cancer by comparing the expressionlevels of one or more BCSM in biological samples from a subject tocontrol samples.

[0009] In a related aspect, the present invention provides a kit fordiagnosing breast cancer. The kit comprises at least one of thefollowing (1) polynucleotide probe that specifically hybridizes to apolynucleotide transcribed from a BCSG, and (2) an antibody capable ofimmunospecific binding to a BCSM.

[0010] In another aspect, the present invention provides apharmaceutical composition for the treatment of breast cancer. Thepharmaceutical composition comprises a pharmaceutically acceptablecarrier and at least one of the following: (1) a BCSM or a functionalvariant of a BCSM, (2) an antibody directed against a BCSM or itsfunctional variant, (3) a vaccine generated using a BCSM or its variant,(4) an agent that modulate an expression level of a BCSG or an activityof a BCSM.

[0011] In a related aspect, the present invention provides a method fortreating breast cancer in a patient with the pharmaceutical compositiondescribed above. The patient may be afflicted with breast cancer, inwhich case the methods provide treatment for the disease. The patientmay also be considered at risk for breast cancer, in which case themethods provide prevention for cancer development.

[0012] In another embodiment, the present invention provides methods forscreening anti-breast cancer agents based on the agents interaction withthe BCSMs, or the agents' effect on the expression of the BCSGs.

[0013] In another embodiment, the present invention provides animalstransgenic for one or more of the BCSGs, or a knockout animal in whichone or more of the BCSGs is disrupted. These animals may be used tostudy the relevance of BCSGs to the development of breast cancer.

[0014] In another embodiment, the present invention provides host cellsharboring a transfected BCSG. These cells may be used for the treatmentof breast cancer.

[0015] Other aspects of the invention will become apparent to theskilled artisan by the following description of the invention.

BRIEF DESCRIPTION OF FIGURES

[0016] The inventions of this application are better understood inconjunction with the following drawings, in which:

[0017]FIG. 1 shows patterns of gene expression in MDA-MB-231 (breastcancer) and MDA/H6 (non-tumorigenic) cell lines. (A) Phosphor images ofgene filters. Five gene filters (gf200, gf201, gf202, gf203, gf211) werehybridized first with radioactively labeled cDNA from MDA-MB-231 cellsand then with that from MDA/H6 cells. (B) Color images derived from thealignment of radioactive images. (C) A scatter plot of expressionintensities of 25,985 genes in MDA-MB-231 and MDA/H6. Each dotrepresents a gene plotted at the coordinate of its two expressionintensities on a log-scale. The genes with the equal intensities arecondensed along a diagonal line. (D) The original and color images of 30genes up-regulated in MDA/H6 with low, medium, and high levels of theexpression. Three equally expressed genes were indicated. Red:up-regulated in MDA/H6; green: down regulated in MDA/H6; yellow: nochanges.

[0018]FIG. 2 shows analysis of images and expression data on thecustomized microarrays. (A and B): The images of two sets of 768 geneson the same glass slide. The image A shows the identical patterns withthe image B. (i and i′): the gene encoding for prostaglandinendoperoxide synthase 2; (ii and ii′) the gene for3-hydroxymethyl-3-methylglutaryl-Coenzyme A lyase; (iii and iii′) thegene for ribosomal protein L10. (C and D) Statistical analysis of theexpression ratios of 202 informative genes between MDA-MB-231 and MDA/H6were detected by two sets of genes (images A and B) on Slide 1 (C) andon Slide 2 (D). (E) The average ratios of the gene expression from Slide1A and 1B were plotted against the average ratios from Slide 2A and 2B.The linear regression and Pearson coefficient of correlation werecomputed from the scatter plots that are on log-scale. The strong linearrelations and high values of Pearson coefficient of correlation (r) areindicated in each comparison. “x”: an gene expression ratio betweenMDA-MB-231 and MDA/H6 on x-axis; “y”: the ratio between these twosamples on the y-axis corresponding to a given “x”.

[0019]FIG. 3 depicts clustering of the gene expression data. (A)Multidimensional scaling analysis. 3-dimentional plot of all 15 cancersamples showing two identical MDA-MB-231 samples (MB231 1 and 2, green),the most dissimilar melanoma sample (MelTis in yellow), three mostsimilar breast cancer samples (BT20, ZR75-1, and BT474 in red) andothers in blue. (B and C) Gene and sample dendrograms from thehierarchical clustering analysis reveal co-regulated genes andrelationship among the samples. Two MDA-MB-231 samples are essentiallyidentical (r=0.982). Human melanoma specimen (MelTis) is the mostdissimilar to MDA-MB-231 (r=0.325). Twelve breast cancer samples areclustered in the center. Three most similar samples were BT20, BT474 andZR-75-1 (r=0.796). The numbers on the nodes indicate the values ofPearson coefficient of correlation. (D) Nine genes with significantlyup-regulated expression (≧2 folds) in at least 10 of 13 breast cancersamples. These nine genes were also over-expressed in the metastaticmelanoma. (E) Ten genes with significantly down-regulated expression(≧0.5 folds) in at least 10 of 13 breast cancer samples. The clone IDand the gene names are listed on the left and the right of the panels,respectively.

[0020]FIG. 4 shows the correlation of thrombomodulin (THBD) RNAexpression to THBD protein expression as measured by cDNA microarraysand Western blots, respectively. (A) The THBD RNA levels in 13 breastcancer cell lines measured by cDNA microarrays using MDA/H6 as thereference. The values of the intensity means (I.M.), the intensitystandard deviations (I.D.), and the calibrated (Cal.) ratios for thetest samples and the reference are the averages derived from the cDNAmicroarray images A and B on each slide (see FIG. 2). The green filledbox and Cal. ratio indicate the decrease of the TH gene in a test samplerelative to the corresponding MDA/H6 reference. (B) Western blot of thewhole cell lysates from the breast cancer cell lines: MDA/H6 (lane 1),MB231 (lane 2), MB436 (lane 3), MB453 (lane 4) and BT549 (lane5), usingthe antibody against THBD (top panel) and the antibody against actin(bottom panel) as a control for loading error. Ninety-eight kilodaltons(kD) and 43 kD indicate the THBD protein and actin protein,respectively. The protein intensities in the lanes 2, 3, 4, and 5approximate the RNA levels in the corresponding breast cancer cells:MB231, MB231, MB436, MB453 and BT549. The lane 1 shows the THBD proteinintensity in the non-tumorigenic breast cancer cell line MDA/H6 thatdisplays the highest RNA level in all the cell lines.

[0021]FIG. 5 show representative images of the pathological sections ofnormal and cancerous breast tissues from Case 1 (A) and Case 6 (B) inTable 6. (A1) A section shows normal breast tissue, of which the mammaryepithelial cells were stained to brown (positive) by the TH antibody(A2). (A3) A tissue section shows infiltrating ductal carcinoma, ofwhich the cancer cells were not stained by the TH antibody (A4). (B1) Asection shows normal mammary epithelial tissue (indicated by thehorizontal arrowheads) and infiltrating ductal carcinoma (indicated bythe vertical arrowheads); (B2) Normal mammary epithelial cells werestained to brown (positive) by the TH antibody; in contrast, the cancercells were not. Magnification: (A1 and A2), 100-fold; (A3 and A4),200-fold; (B1 and B2), 40 fold.

DETAILED DESCRIPTION OF THE INVENTION

[0022] The following detailed description is presented to enable anyperson skilled in the art to make and use the invention. For purposes ofexplanation, specific nomenclature is set forth to provide a thoroughunderstanding of the present invention. However, it will be apparent toone skilled in the art that the specific nomenclature is not required topractice the invention. Descriptions of specific applications areprovided only as representative examples. Various modifications to thepreferred embodiments will be readily apparent to one skilled in theart, and the general principles defined herein may be applied to otherembodiments and applications without departing from the scope of theinvention. The present invention is not intended to be limited to theembodiments shown, but is to be accorded the widest possible scopeconsistent with the principles and features disclosed herein.

[0023] The present invention is generally directed to compositions andmethods for the diagnosis, treatment, and prevention of breast cancer.The present invention is based on the discovery of transcribedpolynucleotides that are either over-expressed or under-expressed inhuman breast cancer cell line MDA-MB-231 as related to thenon-tumorigenic derivative cell line MDA/H6.

[0024] Definitions and Terms

[0025] To facilitate an understanding of the present invention, a numberof terms and phrases are defined below:

[0026] As used herein, the term “breast cancer specific gene (BCSG)”refers to a gene that is over-expressed by at least two-fold (i.e. ≧200%of normal) or under-expressed by at least two-fold (i.e., ≦50% ofnormal) in breast cancer tissue or cell lines relative to normal tissueor cell lines. Specifically, BCSG refers to the genes listed in Table 1and the alleles of these genes.

[0027] As used herein, “a breast cancer-specific marker (BCSM)” refersto a polynucleotide transcribed from a BCSG or a polypeptide translatedfrom such a polynucleotide. BCSM and “BCSG product” are usedinterchangeably.

[0028] As used herein, “a BCSM and its variants” refers to variants of apolynucleotide transcribed from a BCSG and variants of a polypepetideencoded by a BCSG.

[0029] As used herein, the terms “polynucleotide” “nucleic acid” and“oligonucleotide” are used interchangeably, and include polymeric formsof nucleotides of any length, either deoxyribonucleotides orribonucleotides, or analogs thereof. The following are non-limitingexamples of polynucleotides: a gene or gene fragment, exons, introns,messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, DNA, cDNA,genomic DNA, recombinant polynucleotides, branched polynucleotides,plasmids, vectors, isolated DNA of any sequence, isolated RNA of anysequence, nucleic acid probes, and primers.

[0030] As used herein, the terms “variants of a polynucleotide” refersto polynucleotides that, as a result of the degeneracy of the geneticcode, encode the same polypeptide as described herein. Some of thesepolynucleotides bear minimal homology to the nucleotide sequence of anynative gene. Nonetheless, polynucleotides that vary due to differencesin codon usage are specifically contemplated by the present invention. Avariant may contain one or more substitutions, additions, deletionsand/or insertions such that the activity or immunogenicity of theencoded polypeptide is not substantially enhanced or diminished,relative to a native polypeptide.

[0031] Variants of a polynucleotide may also be substantially homologousto a native gene, or a portion or complement thereof. Suchpolynucleotide variants are capable of hybridizing under moderatelystringent conditions to a naturally occurring DNA sequence encoding anative breast tumor protein (or a complementary sequence). Suitablemoderately stringent conditions include prewashing in a solution of5×SSC, 0.5% SDS. 1.0 mM EDTA (pH 8.0); hybridizing at 50° C.-65° C.,5×SSC, overnight; followed by washing twice at 65° C. for 20 minuteswith each of 2×, 0.5× and 0.2×SSC containing 0.1% SDS. Standardhybridization techniques are described in Sambrook et al., MolecularCloning. A Laboratory Manual, Cold Spring Harbor Laboratories, ColdSpring Harbor, N.Y., 1989.

[0032] As used herein, a “variant of a polypeptide” is a polypeptidethat differs from a native polypeptide in one or more substitutions,deletions, additions and/or insertions, such that the functionality ofthe polypeptide is not substantially enhanced or diminished. In otherwords, a variant retains the biological activities of the nativepeptide. The biological activities of the variant may be enhanced ordiminished by less than 50%, preferably less than 20%, relative to thenative polypeptide. Similarly, the ability of a variant to react withantigen-specific antisera may be enhanced or diminished by less than50%, preferably less than 20%, relative to the native polypeptide. Suchvariants may generally be identified by modifying one of the abovepolypeptide sequences and evaluating the reactivity of the modifiedpolypeptide with antigen-specific antibodies or antisera as describedherein.

[0033] Preferably, a variant polypeptide contains conservativesubstitutions. A “conservative substitution” is one in which an aminoacid is substituted for another amino acid that has similar properties,such that one skilled in the art of peptide chemistry would expect thesecondary structure and hydropathic nature of the polypeptide to besubstantially unchanged. Amino acid substitutions may generally be madeon the basis of similarity in polarity, charge, solubility,hydrophobicity, hydrophilicity and/or the amphipathic nature of theresidues. For example, negatively charged amino acids include asparticacid and glutamic acid; positively charged amino acids include lysineand arginine; and amino acids with uncharged polar head groups havingsimilar hydrophilicity values include leucine, isoleucine and valine;glycine and alanine; asparagine and glutamine; and serine, threonine,phenylalanine and tyrosine. Variants may also be modified by, forexample, the deletion or addition of amino acids that have minimalinfluence on the immunogenicity, secondary structure and hydropathicnature of the polypeptide.

[0034] Polypeptide variants preferably exhibit at least about 70%, morepreferably at least about 90% and most preferably at least about 95%homology to the original polypeptide.

[0035] A polypeptide variant also include a polypeptides that ismodified from the original polypeptides by either natural processes,such as posttranslational processing, or by chemical modificationtechniques which are well known in the art. Modifications can occuranywhere in a polypeptide, including the peptide backbone, the aminoacid side-chains and the amino or carboxyl termini. It will beappreciated that the same type of modification may be present in thesame or varying degrees at several sites in a given polypeptide. Also, agiven polypeptide may contain many types of modifications. Polypeptidesmay be branched, for example, as a result of ubiquitination, and theymay be cyclic, with or without branching. Cyclic, branched, and branchedcyclic polypeptides may result from posttranslation natural processes ormay be made by synthetic methods. Modifications include acetylation,acylation, ADP-ribosylation, amidation, covalent attachment of flavin,covalent attachment of a heme moiety, covalent attachment of anucleotide or nucleotide derivative, covalent attachment of a lipid orlipid derivative, covalent attachment of phosphotidylinositol,cross-linking, cyclization, disulfide bond formation, demethylation,formation of covalent cross-links, formation of cysteine, formation ofpyroglutamate, formylation, gamma-carboxylation, glycosylation, GPIanchor formation, hydroxylation, iodination, methylation,myristoylation, oxidation, pegylation, proteolytic processing,phosphorylation, prenylation, racemization, selenoylation, sulfation,transfer-RNA mediated addition of amino acids to proteins such asarginylation, and ubiquitination.

[0036] As used herein, a “biologically active portion” of a polypeptideencoded by a BCSG includes a fragment of the polypeptide comprisingamino acid sequences derived from the original polypeptide, whichinclude fewer amino acids than the full length polypeptide, and exhibitat least one activity of the full length polypeptide. Typically,biologically active portions comprise a domain or motif with at leastone activity of the full length polypeptide. A biologically activeportion of a polypeptide encoded by a BCSG can be a polypeptide whichis, for example, 10, 25, 50, 100, 200 or more amino acids in length.

[0037] As used herein, an “immunologenic portion” or “epitope” of apolypeptide encoded by a BCSG includes a fragment of the originalpolypeptide comprising amino acid sequences sufficiently homologous toor derived from the amino acid sequence of the original polypeptide,which include fewer amino acids than the full length polypeptide and canbe used as an antigen to stimulate anti-BCSG peptide immune response.

[0038] As used herein, the term “modulation” includes, in its variousgrammatical forms (e.g., “modulated”, “modulation”, “modulating”, etc.),up-regulation, induction, stimulation, potentiation, inhibition,down-regulation, or suppression.

[0039] As used herein, the term “control sequences” or “regulatorysequences” refers to DNA sequences necessary for the expression of anoperably linked coding sequence in a particular host organism. The term“control/regulatory sequence” is intended to include promoters,enhancers and other expression control elements (e.g., polyadenylationsignals). Control/regulatory sequences include those which directconstitutive expression of a nucleotide sequence in many types of hostcells and those which direct expression of the nucleotide sequence onlyin certain host cells (e.g., tissue-specific regulatory sequences).

[0040] A nucleic acid sequence is “operably linked” to another nucleicacid sequence when it is placed into a functional relationship withanother nucleic acid sequence. For example, coding sequences of a BCSGcan be operably linked to the regulatory sequences in a manner whichallows for expression of the BCSG (e.g., in an in vitrotranscription/translation system or in a host cell when the vector isintroduced into the host cell). DNA for a presequence or secretoryleader is operably linked to DNA for a polypeptide if it is expressed asa preprotein that participates in the secretion of the polypeptide; apromoter or enhancer is operably linked to a coding sequence if itaffects the transcription of the sequence; or a ribosome binding site isoperably linked to a coding sequence if it is positioned so as tofacilitate translation. Generally, “operably linked” means that the DNAsequences being linked are contiguous, and, in the case of a secretoryleader, contiguous and in reading phase. However, enhancers do not haveto be contiguous. Linking is accomplished by ligation at convenientrestriction sites. If such sites do not exist, the syntheticoligonucleotide adaptors or linkers are used in accordance withconventional practice.

[0041] As used herein, the term “immunospecific binding” refers to thespecific binding of an antibody to an antigen at an affinity that is atleast 10⁵M⁻¹.

[0042] As used herein, the term “biomolecules” refers to moleculeshaving a bioactivity in a mammal. Examples of biomolecules include, butare not limited to, amino acids, nucleic acids, lipids, carbohydrates,polypeptides, polynucleotides, and polysaccsharides.

[0043] Breast Cancer Specific Genes

[0044] Breast cancer consists of extensively heterogeneous tumors andindividual tumor cells may have specific genetic defects that determinegene expression patterns. Identification of expression profiles ofmultiple cancer samples may reveal genes and their expression patternsthat consist of portions specific to the individual samples and commonto most, if not all, samples studied. The common expression patternsmight represent a common “passage” through which the cells evolve fromone status to another. Although the high throughput technology DNAmicroarray is very useful to reveal genome-wide gene expressionprofiles, high density microarrays of thousands of genes are currentlytoo expensive for routine research activities in majority laboratories.

[0045] The present invention uses an alternative approach to combinehigh density gene filters and low-cost high quality microarrays to studygenome-wide gene expression. Gene expression profiles between theparental metastatic breast cancer cell line MDA-MB-231 and thechromosome 6-mediated suppressed non-tumorigenic derivative cell lineMDA/H6 were initially compared using gene filters with 19,592 uniquehuman genes/6,393 controls and radioactive detection technique. Sixhundred and fifty-one genes were found to have more than 800 radioactivesignal intensities and more than 2-fold changes in expression betweenthe parental breast cancer cell line MDA-MB-231 and the non-tumorigeniccell line MDA/H6.

[0046] The 651 differentially expressed genes were further examinedusing customized DNA microarrays and fluorescence detection techniques.Since gene expression levelsin the same cells detected by microarrayscan be affected by many factors including cell culture conditions, RNApurification, cDNA labeling methods and the quality of microarrays, highquality microarrays were used in the present invention to reduce thevariance that could otherwise be introduced by different microarrayslides. Strong positive linear relations with high values of Pearsoncoefficient of correlation were obtained between 2 sets of genes on thesame slides and between the genes on the different slides, demonstratingthe consistency of the microarrays and reproducibility of theexperiments. The microarray analysis revealed 202 genes that wereexpressed differentially in breast cancer cell lines (n=10) and clinicalbreast cancer specimens (n=3) as related to normal tissues. The genesidentified by the microarray and their expression profiles are listed inTables 1 and 2, respectively. TABLE 1 Genes with informative expressionprofiles in breast cancer cell lines Clone ID Gene Name Title PlatePosition  23185 TNC hexabrachion (tenascin C, cytotactin) LCC9d11  23831ALDOC aldolase C, fructose-bisphosphate LCC1e11  26617 ALCAM activatedleucocyte cell adhesion molecule LCC2b1  26711 NCBP2 nuclear cap bindingprotein subunit 2, 20 kD LCC1g10  28098 LOC57862 clones 23667 and 23775zinc finger protein LCC1e5  28116 karyopherin a2 karyopherin alpha 2(RAG cohort 1, importin alpha 1) LCC9e1  30476 ESTs ESTs LCC9d8  32517FLJ10509 hypothetical protein FLJ10509 LCC8e12  33949 PRPSAP1phosphoribosyl pyrophosphate synthetase-associated protein 1 LCC1d8 36191 Fibronectin 1 fibronectin 1 LCC9d10  39884 IMPDH1 IMP (inosinemonophosphate) dehydrogenase 1 LCC8a8  40026 SLC25A4 solute carrierfamily 25 (mitochondrial carrier; adenine nucleotide translocator),member 4 LCC8g5  44178 TEGT testis enhanced gene transcript LCC9d7 44255 RPML3 ribosomal protein, mitochondrial, L3 LCC8a7  45641 MAP2K3mitogen-activated protein kinase kinase 3 LCC3a10  45801 ESTs ESTsLCC5b5  49496 AMID programmed cell death 8 (apoptosis-inducing factor)LCC9a12  49553 ARF4L ADP-ribosylation factor 4-like LCC8g6  49987 GRIA2glutamate receptor, ionotropic, AMPA 2 LCC8e6  51718 ESTs ESTs LCC9b3 66686 RPL10 ribosomal protein L10 LCC1a11  71101 PROCR protein Creceptor, endothelial (EPCR) LCC2h2  79710 KIAA0174 KIAA0174 geneproduct LCC2e10  80910 SLC1A5 solute carrier family 1 (neutral aminoacid transporter), member 5 LCC2e8  108667 SF3A1 splicing factor 3a,subunit 1, 120 kD LCC8b6  112576 ESTs ESTs LCC3e1  114101 ESTs ESTsLCC9c8  127519 POH1 26S proteasome-associated pad1 homolog LCC1f11 127821 ACP5 acid phosphatase 5, tartrate resistant LCC2a8  128243 ADKadenosine kinase LCC2b5  129585 EST(Metallothionein2) EST, Moderatelysimilar to Cd-7 Metallothionein-2 [H. sapiens] LCC3d9  131563 FLJ13443Homo sapiens cDNA FLJ13443 fis, clone PLACE1002853 LCC4a1  134495FLJ10976 Homo sapiens cDNA FLJ10976 fis, clone PLACE1001399 LCC4a10 135083 GRP58 glucose regulated protein, 58 kD LCC8c10  136798Fibronectin 1 fibronectin 1 LCC9a5  138345 PTP IVA protein tyrosinephosphatase type IVA, member 1 LCC9a6  139883 ESTs ESTs LCC4b2  142586MCT-1 MCT-1 protein LCC4f6  144926 ESTs ESTs, Weakly similar to B0495.6[C. elegans] LCC3e5  147050 PTGS2 prostaglandin-endoperoxide synthase 2(prostaglandin G/H synthase and cyclooxygenase) LCC9d4  147338 ESTs ESTsLCC9a7  163097 MAM melanoma adhesion molecule LCC9d1  173554 SFRS3splicing factor, arginine/serine-rich 3 LCC6d8  191603 TUBB tubulin,beta polypeptide LCC8a4  198871 ESTs ESTs LCC9b6  201436 LCC9c4  205185THBD thrombomodulin LCC1a7  207358 SLC2A1 solute carrier family 2(facilitated glucose transporter), member 1 LCC3b3  208001 CD59 CD59antigen p18-20 (antigen identified by monoclonal antibodies 16.3A5,EJ16, EJ30, EL32 LCC2b4 and G344)  208699 ESTs ESTs LCC4f7  212165 PRDX2peroxiredoxin 2 LCC2h7  220376 ESTs ESTs LCC9b8  221632 EIF2B2eukaryotic translation initiation factor 2B, subunit 2 (beta, 39 kD)LCC6e1  223141 ESTs ESTs LCC9d9  232772 EST(Metallothionein-1B ESTs,Highly similar to MT1B_HUMAN METALLOTHIONEIN-1B [H. sapiens] LCC1c12 233581 HIP2 huntingtin interacting protein 2 LCC3b10  234398TCCCIA00427 Homo sapiens clone TCCCIA00427 mRNA sequence LCC3g11  236305HARS histidyl-tRNA synthetase LCC8c12  239877 HDAC3 histone deacetylase3 LCC3b9  244147 ZFP92 zinc finger protein homologous to Zfp92 in mouseLCC3e6  245547 KIAA0700 KIAA0700 protein LCC6c4  251753 ESTs ESTs LCC5c9 257197 NRBF-2 nuclear receptor binding factor-2 LCC4h8  271478MAX-interacting protein MAX-interacting protein 1 LCC9b10  276547 DNMT1DNA (cytosine-5-)-methyltransferase 1 LCC8b11  284592 PRO1659 PRO1659protein LCC4f8  292213 PERQ1 PERQ amino acid rich, with GYF domain 1LCC1c7  295140 FLJ0330 hypothetical protein FLJ10330 LCC4d2  295410 ESTsESTs LCC3f6  296998 ART4 ADP-ribosyltransferase 4 LCC1h9  298155 ACADMacyl-Coenzyme A dehydrogenase, C-4 to C-12 straight chain LCC2b2  298965COX6B cytochrome c oxidase subunit VIb LCC5g11  307532 EIF4A2 eukaryotictranslation initiation factor 4A, isoform 2 LCC8d7  310493 FACL3fatty-acid-Coenzyme A ligase, long-chain 3 LCC3c10  321189 RAP1B RAP1B,member of RAS oncogene family LCC3d12  321661 PPP2R5C proteinphosphatase 2, regulatory subunit B (B56), gamma isoform LCC1b4  321859ESTs ESTs LCC4h10  322759 SNAPC5 small nuclear RNA activating complex,polypeptide 5, 19 kD LCC4b8  323474 ARF1 ADP-ribosylation factor 1LCC8d11  325062 SLC20A1 solute carrier family 20 (phosphatetransporter), member 1 LCC1e3  325102 EST(CTB2) ESTs, Moderately similarto CTB2_HUMAN C-TERMINAL BINDING PROTEIN 2□ LCC3d10 [H. sapiens]  327304H326 H326 LCC1f9  340840 FLJ20263 (AKAP450) Homo sapiens cDNA FLJ20263fis, clone COLF7804, highly similar to AJ131693 LCC3f3 Homo sapiens mRNAfor AKAP450 protein  342378 DUSP5 dual specificity phosphatase 5 LCC1d5 346009 PFKL phosphofructokinase, liver LCC8f9  358531 JUN v-jun aviansarcoma virus 17 oncogene homolog LCC3b4  359835 SAT spermidine/spermineN1-acetyltransferase LCC8a5  359933 GNAS1 guanine nucleotide bindingprotein (G protein), alpha stimulating activity polypeptide 1 LCC8d9 361565 GLUD1 glutamate dehydrogenase 1 LCC8a11  365930 TAF2F TATA boxbinding protein (TBP)-associated factor, RNA polymerase II, F, 55 kDLCC1e12  399562 NUP54 nucleoporin p54 LCC6h2  430318 PVALB parvalbuminLCC2c1  436051 ESTs ESTs, Weakly similar to putative p150 [H. sapiens]LCC6h10  449112 EST(G3PDH) ESTs, Highly similar to G3P2_HUMANGLYCERALDEHYDE 3-PHOSPHATE LCC6h9 DEHYDROGENASE, LIVER□ [H. sapiens] 454970 DKFZP434G032 DKFZP434G032 protein LCC9g12  469151 EIF2S2eukaryotic translation initiation factor 2, subunit 2 (beta, 38 kD)LCC8f10  471863 DKFZp586C1817 Homo sapiens mRNA; cDNA DKFZp586C1817(from clone DKFZp586C1817) LCC9h9  509516 LOC56966 hypothetical proteinfrom EUROIMAGE 1034327 LCC5c5  511521 CANX calnexin LCC2a6  511586HNRPA1 heterogeneous nuclear ribonucleoprotein A1 LCC8c11  564803 FOXM1forkhead box M1 LCC2h3  628357 ACTN3 actinin, alpha 3 LCC2a11  665774EIF4E eukaryotic translation initiation factor 4E LCC1h7  711959 RPC62polymerase (RNA) III (DNA directed) (62 kD) LCC2f12  712840 STAT5Bsignal transducer and activator of transcription 5B LCC2d4  712848 MADDMAP-kinase activating death domain LCC2h4  713647 TSPAN-3 tetraspan 3LCC2f6  714210 RY1 putative nucleic acid binding protein RY-1 LCC3a4 725274 TTC1 tetratricopeptide repeat domain 1 LCC2d3  730149 TCEA2transcription elongation factor A (SII), 2 LCC1d4  739183 CD68 CD68antigen LCC3a12  739625 KIAA0973 KIAA0973 protein LCC2h10  739993 BREbrain and reproductive organ-expressed (TNFRSF1A modulator) LCC2g11 740914 CTBP1 C-terminal binding protein 1 LCC2h5  741067 SMARCD2SWI/SNF related, matrix associated, actin dependent regulator ofchromatin, subfamily d, LCC1f6 member 2  741988 ACY1 aminoacylase 1LCC8f6  745604 BCAR1 breast cancer anti-estrogen resistance 1 LCC8g8 753313 LAPTM5 Lysosomal-associated multispanning membrane protein-5LCC1e2  753457 NDUFS1 NADH dehydrogenase (ubiquinone) Fe-S protein 1 (75kD) (NADH-coenzyme Q reductase) LCC1b11  753897 AMFR autocrine motilityfactor receptor LCC2a10  755444 TMSB4X thymosin, beta 4, X chromosomeLCC6e12  756490 BCAT2 branched chain aminotransferase 2, mitochondrialLCC5a12  756600 PPIB peptidylprolyl isomerase B (cyclophilin B) LCC8d8 756769 CHAF1B chromarin assembly factor 1, subunit B (p60) LCC8b3 756968 EFNB1 ephrin-B1 LCC2g7  758365 OS4 conserved gene amplified inosteosarcoma LCC3b7  758662 PSMD9 proteasome (prosome, macropain) 26Ssubunit, non-ATPase, 9 LCC2e1  759200 DHPS deoxyhypusine synthase LCC8e9 760298 PRSC1 protease, cysteine, 1 (legumain) LCC2e7  770080 PXNpaxillin LCC1d2  770388 CLDN4 claudin 4 LCC5b1  773147 FLJ10491 Homosapiens cDNA FLJ10491 fis, clone NT2RP2000239 LCC5e3  773367 COMTcatechol-O-methyltransferase LCC8f4  774071 CLTH Clathrin assemblylymphoid-myeloid leukemia gene LCC2g2  781704 TRIP7 thyroid hormonereceptor interactor 7 LCC2g12  783698 KIAA0188 KIAA0188 protein LCC2e12 784278 SF100 nuclear antigen Sp100 LCC2c9  784841 EIF2S3 eukaryotictranslation initiation factor 2, subunit 3 (gamma, 52 kD) LCC2b10 786048 E2F4 E2F transcription factor 4, p107/p130-binding LCC3a3 788574 GCN5L2 GCN5 (general control of amino-acid synthesis, yeast,homolog)-like 2 LCC2g8  789232 PSMD4 proteasome (prosome, macropain) 26Ssubunit, non-ATPase, 4 LCC2g4  795282 HSPC126 HSPC126 protein LCC4h3 795330 NR1D1 nuclear receptor subfamily 1, group D, member 1 LCC2b11 795888 RBBP2 retinoblastoma-binding protein 2 LCC2b12  809517 PRO2605hypothetical protein PRO2605 LCC4g7  809648 ZNF162 zinc finger protein162 LCC2g10  809835 HNRPC heterogeneous nuclear ribonucleoprotein C(C1/C2) LCC8d12  809992 PSMD2 proteasome (prosome, macropain) 26Ssubunit, non-ATPase, 2 LCC1g9  809992 PSMD2 proteasome (prosome,macropain) 26S subunit, non-ATPase, 2 LCC8b8  810019 HNRPD heterogeneousnuclear ribonucleoprotein D (AU-rich element RNA-binding protein 1, 37kD) LCC8a10  810791 MNAT1 menage a trois 1 (CAK assembly factor) LCC8b7 810873 SCNN1A sodium channel, nonvoltage-gated 1 alpha LCC1a8  811792GSS glutathione synthetase LCC1h2  813158 DRG2 developmentally regulatedGTP-binding protein 2 LCC1g11  813280 ADSL adenylosuccinate lyaseLCC2a12  813426 G53955 GS3955 protein LCC1f4  813648 DLDdihydrolipoamide dehydrogenase (E3 component of pyruvate dehydrogenasecomplex, LCC8b10 2-oxo-glutarate complex, branched chain keto aciddehydrogenase complex)  813742 PTK7 PTK7 protein tyrosine kinase 7LCC1b2  814508 PPP1R7 protein phosphatase 1, regulatory subunit 7 LCC2h9 814595 PRKCBP1 protein kinase C binding protein 1 LCC2d5  814636SMARCA2 SWI/SNF related, matrix associated, actin dependent regulator ofchromatin, subfamily a, LCC2e3 member 2  815542 MX1 myxovirus(influenza) resistance 1, homolog of marine (interferon-inducibleprotein p78) LCC2c10  815575 ACTR1A ARP1 (actin-related protein 1,yeast) homolog A (centractin alpha) LCC8f3  823930 ARPC1A actin relatedprotein 2/3 complex, subunit 1A (41 kD) LCC1g7  824024 NQO2 NAD(P)Hmenadione oxidoreductase 2, dioxin-inducible LCC2c3  824031 HSJ2 heatshock protein, DNAJ-like 2 LCC3a7  824602 IFI16 interferon,gamma-inducible protein 16 LCC2f7  825470 TOP2A topoisomerase (DNA) IIalpha (170 kD) LCC2b7  838366 HMGCL3-hydroxymethyl-3-methylglutaryl-Coenzyme A lyase(hydroxymethylglutaricaciduria) LCC8g4  840404 MGAT2 mannosyl(alpha-1,6-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferaseLCC2g6  840940 PABPC1 poly(A)-binding protein, cytoplasmic 1 LCC2c8 841691 MNPEP methionine aminopeptidase; eIF-2-associated p67 LCC8c9 843016 P130 nucleolar phosphoprotein p130 LCC2f5  843328 DUSP12 dualspecificity phosphatase 12 LCC5c2  852520 UQCRC2 ubiquinol-cytochrome creductase core protein II LCC8e2  853570 SLC25A6 solute carrier family25 (mitochondrial carrier; adenine nucleotide translocator), member 6LCC8f5  855910 LGALS3 lectin, galactoside-binding, soluble, 3 (galectin3) LCC5a9  866882 FDFT1 farnesyl-diphosphate farnesyltransferase 1LCC8e8  868368 TMSB4X thymosin, beta 4, X chromosome LCC5a11  877613DCTN1 dynactin 1 (p150, Glued (Drosophila) homolog) LCC2h8  877832DXS1357E accessory proteins BAP31/BAP29 LCC8e5  878545 RPL18 ribosomalprotein L18 LCC6c9  884644 HBG1 hemoglobin, gamma A LCC5a10  897164CTNNA1 catenin (cadherin-associated protein), alpha 1 (102 kD) LCC8e7 897177 PGAM1 phosphoglycerate mutase 1 (brain) LCC8e3  897626 PRO2706hypothetical protein PRO2706 LCC2h11  897880 CCT4 chaperonin containingTCP1, subunit 4 (delta) LCC8d6  897983 KIAA0106 anti-oxidant protein 2(non-selenium glutathione peroxidase, acidic calcium-independent LCC2f9phospholipase A2)  898262 UBE1 ubiquitin-activating enzyme E1 (A1S9T andBN75 temperature sensitivity complementing) LCC8c3  949928 ZNF220 zincfinger protein 220 LCC2e2  950489 SOD1 superoxide dismutase 1, soluble(amyotrophic lateral sclerosis 1 (adult)) LCC8b12  950682 PFKPphosphofructokinase, platelet LCC8c5  951117 SHMT2 serinehydroxymethyltransferase 2 (mitochondrial) LCC3b6  951313 GP1 glucosephosphate isomerase LCC5c6  969854 CALM3 calmodulin 3 (phosphorylasekinase, delta) LCC8e4  971367 RPS8 ribosomal protein S8 LCC6c10 1160558PTS 6-pyruvoyltetrahydropterin synthase LCC6c3 1340595 HNRPLheterogeneous nuclear ribonucleoprotein L LCC6b12 1416782 CKB creatinekinase, brain LCC8f7 1473300 HADHA hydroxyacyl-Coenzyme Adehydrogenase/3-ketoacyl-Coenzyme A thiolase/enoyl-Coenzyme A LCC8f11hydratase (trifunctional protein), alpha subunit 1475028 RPS27 ribosomalprotein S27 (metallopanstimulin 1) LCC6c8 1475730 CCT6A chaperonincontaining TCP1, subunit 6A (zeta 1) LCC8f12

[0047] TABLE 2 Gene expression profiles in breast cancer cell lines GeneName Clone ID Plate Pos. MB231-1A MB231-1B MB231-2A MB231-2B MB231-1 TNC 23185 LCC9d11 0.744 0.811 0.773 0.611 0.7775 ALDOC  23831 LCC1e11 0.8770.944 1.038 1.043 0.9105 ALCAM  26617 LCC2b1 0.563 0.595 0.582 0.5260.579 NCBP2  26711 LCC1g10 0.55 0.562 0.603 0.615 0.556 LOC57862  28098LCC1e5 0.846 0.849 0.713 0.688 0.8475 karyopherin a2  28116 LCC9e1 1.1671.23 1.169 1.204 1.1985 ESTs  30476 LCC9d8 2.033 2.229 2.179 1.895 2.131FLJ10509  32517 LCC8e12 1.039 1.09 1.14 1.028 1.0645 PRPSAP1  33949LCC1d8 1.38 1.575 1.347 1.296 1.4775 Fibronectin 1  36191 LCC9d10 0.5590.59 0.639 0.542 0.5745 IMPDH1  39884 LCC8a8 0.541 0.572 0.547 0.530.5565 SLC25A4  40026 LCC8g5 1.716 1.829 1.465 1.704 1.7725 TEGT  44178LCC9d7 1.4 1.458 1.511 1.291 1.429 RPML3  44255 LCC8a7 0.594 0.627 0.5520.639 0.6105 MAP2K3  45641 LCC3a10 0.605 0.627 0.602 0.67 0.616 ESTs 45801 LCC5b5 0.62 0.66 0.661 0.579 0.64 AMID  49496 LCC9a12 1.494 1.7091.686 1.712 1.6015 ARF4L  49553 LCC8g6 1.379 1.483 1.443 1.321 1.431GRIA2  49987 LCC8e6 0.803 0.919 0.796 0.74 0.861 ESTs  51718 LCC9b3 1.881.979 2.247 1.845 1.9295 RPL10  66686 LCC1a11 2.54 2.746 2.748 2.7842.643 PROCR  71101 LCC2h2 0.905 0.765 0.77 0.769 0.835 KIAA0174  79710LCC2e10 0.963 1.036 1.137 1.114 0.9995 SLC1A5  80910 LCC2e8 3.853 4.2133.84 4.022 4.033 SF3A1  108667 LCC8b6 0.574 0.585 0.695 0.498 0.5795ESTs  112576 LCC3e1 0.684 0.75 0.973 0.699 0.717 ESTs  114101 LCC9c80.685 0.716 0.682 0.733 0.7005 POH1  127519 LCC1f11 1.143 1.191 1.1551.102 1.167 ACP5  127821 LCC2a8 0.723 0.775 0.866 0.785 0.749 ADK 128243 LCC2b5 0.506 0.801 0.854 0.803 0.6535 EST  129585 LCC3d9 0.8580.879 0.868 0.863 0.8685 FLJ13443  131563 LCC4a1 0.844 0.87 0.83 0.8250.857 FLJ10976  134495 LCC4a10 0.851 0.878 0.947 0.896 0.8645 GRP58 135083 LCC8c10 0.651 0.67 0.612 0.602 0.6605 Fibronectin 1  136798LCC9a5 0.548 0.574 0.559 0.557 0.561 PTP IVA  138345 LCC9a6 0.405 0.4060.418 0.426 0.4055 ESTs  139883 LCC4b2 0.679 0.762 0.742 0.691 0.7205MCT-1  142586 LCC4f6 1.36 1.416 1.389 1.488 1.388 ESTs  144926 LCC3e50.859 0.988 0.923 1.047 0.9235 PTGS2  147050 LCC9d4 0.066 0.088 0.060.065 0.077 ESTs  147338 LCC9a7 0.67 0.737 0.71 0.769 0.7035 MAM  163097LCC9d1 0.652 0.839 0.681 0.634 0.7455 SFRS3  173554 LCC6d8 1.796 2.0761.532 1.884 1.936 TUBB  191603 LCC8a4 0.892 0.96 0.871 0.951 0.926 ESTs 198871 LCC9b6 0.99 1.022 1.053 0.904 1.006  201436 LCC9c4 1.138 1.281.219 1.231 1.209 THBD  205185 LCC1a7 0.173 0.185 0.114 0.108 0.179SLC2A1  207358 LCC3b3 0.593 0.592 0.649 0.696 0.5925 CD59  208001 LCC2b40.818 0.914 0.832 0.898 0.866 ESTs  208699 LCC4f7 0.574 0.641 0.6710.596 0.6075 PRDX2  212165 LCC2h7 0.735 0.782 0.721 0.721 0.7585 ESTs 220376 LCC9b8 1.517 1.46 1.46 1.283 1.4885 EIF2B2  221632 LCC6e1 0.4430.474 0.43 0.499 0.4585 ESTs  223141 LCC9d9 1.125 1.116 1.18 0.9621.1205 EST(MTT-1B)  232772 LCC1c12 0.84 0.848 1.174 0.92 0.844 HIP2 233581 LCC3b10 0.615 0.643 0.623 0.59 0.629 TCCCIA00427  234398 LCC3g111.221 1.317 1.198 1.333 1.269 HARS  236305 LCC8c12 1.073 1.131 1.2391.175 1.102 HDAC3  239877 LCC3b9 1.046 1.013 1.062 1.175 1.0295 ZFP92 244147 LCC3e6 0.829 0.865 0.784 0.89 0.847 KIAA0700  245547 LCC6c40.646 0.713 0.668 0.769 0.6795 ESTs  251753 LCC5c9 0.852 0.901 0.8330.878 0.8765 NRBF-2  257197 LCC4h8 0.981 0.963 0.908 0.967 0.972 MAX-IP1 271478 LCC9b10 0.758 0.789 0.772 0.687 0.7735 DNMT1  276547 LCC8b110.923 1.099 1.13 1.059 1.011 PRO1659  284592 LCC4f8 1.06 1.145 1.0121.042 1.1025 PERQ1  292213 LCC1c7 1.213 1.347 1.097 1.096 1.28 FLJ10330 295140 LCC4d2 1.248 1.565 1.34 1.17 1.4065 ESTs  295410 LCC3f6 1.3621.364 1.409 1.355 1.363 ART4  296998 LCC1h9 0.697 0.762 0.602 0.6060.7295 ACADM  298155 LCC2b2 1.56 1.724 1.612 1.633 1.642 COX6B  298965LCC5g11 1.604 1.678 1.565 1.799 1.641 EIF4A2  307532 LCC8d7 1.494 2.1591.418 1.37 1.8265 FACL3  310493 LCC3c10 0.884 1.289 1.328 1.478 1.0865RAP1B  321189 LCC3d12 1.156 1.294 1.227 1.057 1.225 PPP2R5C  321661LCC1b4 0.696 0.734 0.768 0.645 0.715 ESTs  321859 LCC4h10 0.471 0.5350.569 0.503 0.503 SNAPC5  322759 LCC4b8 0.94 0.994 0.952 0.978 0.967ARF1  323474 LCC8d11 1.149 1.114 1.09 0.95 1.1315 SLC20A1  325062 LCC1e31.269 1.438 1.257 1.385 1.3535 EST(CTB2)  325102 LCC3d10 1.258 1.2871.149 1.201 1.2725 H326  327304 LCC1f9 0.833 1.007 0.865 0.929 0.92FLJ20263(AKAP450)  340840 LCC3f3 0.883 1.089 0.803 0.673 0.986 DUSP5 342378 LCC1d5 0.45 0.488 0.429 0.478 0.469 PFKL  346009 LCC8f9 1.1321.188 1.099 1.027 1.16 JUN  358531 LCC3b4 0.527 0.565 0.523 0.472 0.546SAT  359835 LCC8a5 0.469 0.517 0.467 0.515 0.493 GNAS1  359933 LCC8d90.929 1.131 0.959 0.825 1.03 GLUD1  361565 LCC8a1 1.114 1.173 1.1821.244 1.1435 TAF2F  365930 LCC1e12 0.888 0.918 0.941 0.992 0.903 NUP54 399562 LCC6h2 1.101 1.05 1.225 1.062 1.0755 PVALB  430318 LCC2c1 0.8660.676 0.796 0.81 0.771 ESTs  436051 LCC6h10 0.712 0.931 0.921 0.9190.8215 EST(G3PDH)  449112 LCC6h9 0.935 0.925 1.034 0.89 0.93DKFZP434G032  454970 LCC9g12 0.724 0.681 0.727 0.67 0.7025 EIF2S2 469151 LCC8f10 1.07 1.08 1.209 0.934 1.075 DKFZp586C1817  471863 LCC9h91.486 1.689 1.51 1.086 1.5875 LOC56966  509516 LCC5c5 1.035 1.219 0.9080.93 1.127 CANX  511521 LCC2a6 1.473 1.601 1.737 1.655 1.537 HNRPA1 511586 LCC8c11 1.492 1.527 1.535 1.566 1.5095 FOXM1  564803 LCC2h30.813 0.884 0.893 0.815 0.8485 ACTN3  628357 LCC2a11 0.782 0.756 0.8210.855 0.769 EIF4E  665774 LCC1h7 0.705 0.752 0.672 0.699 0.7285 RPC62 711959 LCC2f12 1.993 2.185 1.827 2.177 2.089 STAT5B  712840 LCC2d40.867 0.91 0.837 0.914 0.8885 MADD  712848 LCC2h4 0.775 0.832 0.8930.854 0.8035 TSPAN-3  713647 LCC2f6 0.715 0.749 0.747 0.784 0.732 RY1 714210 LCC3a4 0.838 0.917 0.832 0.906 0.8775 TTC1  725274 LCC2d3 0.960.965 0.976 0.955 0.9625 TCEA2  730149 LCC1d4 0.796 1.113 1.051 1.030.9545 CD68  739183 LCC3a12 0.88 0.92 0.974 0.952 0.9 KIAA0973  739625LCC2h10 1.495 1.761 1.496 1.612 1.628 BRE  739993 LCC2g11 0.926 0.9660.963 1.082 0.946 CTBP1  740914 LCC2h5 1.039 1.061 1.103 1.064 1.05SMARCD2  741067 LCC1f6 1.212 1.206 1.103 1.037 1.209 ACY1  741988 LCC8f61.556 1.691 1.71 1.413 1.6235 BCAR1  745604 LCC8g8 0.698 0.78 0.7290.718 0.739 LAPTM5  753313 LCC1e2 0.845 0.75 0.762 0.815 0.7975 NDUFS1 753457 LCC1b11 0.997 1.149 1.107 1.057 1.073 AMFR  753897 LCC2a10 1.5161.667 1.592 1.535 1.5915 TMSB4X  755444 LCC6e12 1.373 1.534 1.192 1.2651.4535 BCAT2  756490 LCC5a12 1.264 1.367 1.235 1.102 1.3155 PPIB  756600LCC8d8 1.434 1.485 1.461 1.31 1.4595 CHAF1B  756769 LCC8b3 0.972 0.9020.891 0.769 0.937 EFNB1  756068 LCC2g7 1.258 1.292 1.17 1.287 1.275 OS4 758365 LCC3b7 1.23 1.278 1.202 1.323 1.254 PSMD9  758662 LCC2e1 0.8220.842 0.826 0.931 0.832 DHPS  759200 LCC8e9 1.048 1.228 1.05 1.161 1.138PRSC1  760298 LCC2e7 0.405 0.422 0.375 0.422 0.4135 PXN  770080 LCC1d20.844 0.852 0.832 0.868 0.848 CLDN4  770388 LCC5b1 5.1407 5.7961 6.2765.436 5.4684 FLJ10491  773147 LCC5e3 1.42 1.466 1.521 1.42 1.443 COMT 773367 LCC8f4 1.074 1.195 1.173 0.939 1.1345 CLTH  774071 LCC2g2 0.7730.811 0.731 0.729 0.792 TRIP7  781704 LCC2g12 0.846 0.937 1.027 0.9920.8915 KIAA0188  783698 LCC2e12 1.417 1.489 1.562 1.435 1.453 SP100 784278 LCC2c9 0.773 0.811 0.803 0.851 0.792 EIF2S3  784841 LCC2b102.165 2.741 2.062 2.676 2.453 E2F4  786048 LCC3a3 1.269 1.422 1.1031.283 1.3455 GCN5L2  788574 LCC2g8 0.601 0.571 0.682 0.634 0.586 PSMD4 789232 LCC2g4 0.996 1.072 1.143 1.174 1.034 HSPC126  795282 LCC4h31.172 1.343 1.487 1.315 1.2575 NR1D1  795330 LCC2b11 0.301 0.363 0.3150.282 0.332 RBBP2  795888 LCC2b12 1.416 1.68 1.423 1.535 1.548 PRO2605 809517 LCC4g7 0.937 0.976 0.894 0.908 0.9565 ZNF162  809648 LCC2g101.518 1.725 1.648 1.797 1.6215 HNRPC  809835 LCC8d12 1.04 1.115 1.3191.011 1.0775 PSMD2  809992 LCC1g9 0.564 0.581 0.634 0.587 0.5725 PSMD2 809992 LCC8b8 0.542 0.573 0.597 0.507 0.5575 HNRPD  810019 LCC8a100.665 0.705 0.73 0.684 0.685 MNAT1  810791 LCC8b7 0.578 0.607 0.6830.591 0.5925 SCNN1A  810873 LCC1a8 1.945 1.789 1.741 1.749 1.867 GSS 811792 LCC1h2 0.366 0.368 0.392 0.366 0.367 DRG2  813158 LCC1g11 0.6020.631 0.615 0.631 0.6165 ADSL  813280 LCC2a12 1.341 1.449 1.439 1.5561.395 GS3955  813426 LCC1f4 1.088 1.165 1.162 1.005 1.1265 DLD  813648LCC8b10 0.703 0.762 0.78 0.599 0.7325 PTK7  813742 LCC1b2 0.509 0.5760.425 0.481 0.5425 PPP1R7  814508 LCC2h9 1.66 1.609 1.458 1.501 1.6345PRKCBP1  814595 LCC2d5 0.754 0.834 0.762 0.686 0.794 SMARCA2  814636LCC2e3 1.96 2.11 2.035 2.137 2.035 MX1  815542 LCC2c10 1.237 1.526 1.1681.083 1.3815 ACTR1A  815575 LCC8f3 1.318 1.384 1.193 1.145 1.351 ARPC1A 823930 LCC1g7 0.876 0.88 0.902 0.818 0.878 NQO2  824024 LCC2c3 0.920.984 0.952 1.186 0.952 HSJ2  824031 LCC3a7 1.182 1.255 1.108 1.1761.2185 IFI16  824602 LCC2f7 0.607 0.613 0.596 0.527 0.61 TOP2A  825470LCC2b7 0.68 0.69 0.671 0.65 0.685 HMGCL  838366 LCC8g4 0.934 0.932 0.8970.951 0.933 MGAT2  840404 LCC2g6 1.537 1.567 1.492 1.521 1.552 PABPC1 840940 LCC2c8 0.56 0.664 0.633 0.586 0.612 MNPEP  841691 LCC8c9 1.0811.135 1.186 1.297 1.108 P130  843016 LCC2f5 1.017 1.067 1.09 0.996 1.042DUSP12  843328 LCC5c2 1.156 1.218 1.183 1.208 1.187 UQCRC2  852520LCC8e2 1.073 1.134 1.116 1.082 1.1035 SLC25A6  853570 LCC8f5 1.724 2.0271.775 1.649 1.8735 LGALS3  855910 LCC5a9 0.741 0.805 0.706 0.772 0.773FSFT1  866882 LCC8e8 0.749 0.779 0.834 0.783 0.764 TMSB4X  868368LCC5a11 1.407 1.484 1.239 1.237 1.4433 DCTN1  877613 LCC2h8 0.878 0.9440.877 0.956 0.911 DXS1357E  877832 LCC8e5 1.342 1.426 1.156 1.283 1.384RPL18  878545 LCC6c9 1.91 2.026 2.03 2.26 1.968 HBG1  884644 LCC5a101.237 1.444 1.376 1.44 1.3405 CTNNA1  897164 LCC8e7 1.061 1.128 0.9481.098 1.0945 PGAM1  897177 LCC8e3 0.993 1.113 0.97 1.1 1.053 PRO2706 897626 LCC2h11 1.06 1.123 1.119 1.013 1.0915 CCT4  897880 LCC8d6 1.0120.997 1.109 0.906 1.0045 KIAA0106  897983 LCC2f9 1.302 1.27 1.26 1.2261.296 UBE1  898262 LCC8c3 0.859 1.01 0.783 0.913 0.9345 ZNF220  949928LCC2e2 1.124 1.114 1.246 1.285 1.119 SOD1  950489 LCC8b12 1.219 1.2551.434 1.124 1.237 PFKP  950682 LCC8c5 1.325 1.398 1.271 1.454 1.3615SHMT2  951117 LCC3b6 2.854 3.233 2.71 2.87 3.0435 GPI  951313 LCC5c61.078 1.16 1.134 1.186 1.119 CALM3  969854 LCC8e4 1.075 1.153 1.0311.042 1.114 RPS8  971367 LCC6c10 1.691 1.806 1.775 1.977 1.7485 PTS1160558 LCC6c3 1.199 1.259 1.163 1.337 1.229 HNRPL 1340595 LCC6b12 0.870.943 0.978 0.948 0.9065 CKB 1416782 LCC8f7 0.23 0.225 0.257 0.2470.2275 HADHA 1473300 LCC8f11 1.113 1.151 1.234 1.041 1.132 RPS27 1475028LCC6c8 1.125 1.225 1.17 1.208 1.175 CCT6A 1475730 LCC8f12 1.472 1.5311.582 1.287 1.5015 Gene Name MB231-2 MelTis BCTis-1 BCTis-2 MB468 ZR75-1BT549 TNC 0.692 3.4165 0.322 0.401 0.083 0.019 0.225 ALDOC 1.0405 3.56850.7555 2.9265 0.19 0.8445 0.75 ALCAM 0.554 2.2295 0.3495 1.2215 0.2580.394 3.323 NCBP2 0.609 1.118 0.498 0.7145 0.412 0.535 0.3855 LOC578620.7005 2.5825 0.899 2.1045 0.9105 1.349 1.0285 karyopherin a2 1.18650.562 0.182 0.4275 1.1675 1.706 0.737 ESTs 2.037 2.8725 0.845 2.07151.45 0.577 0.634 FLJ10509 1.084 1.567 0.5135 1.509 1.2675 1.106 0.9395PRPSAP1 1.3215 3.6375 1.1135 2.146 1.6545 2.5855 0.9365 Fibronectin 10.5905 1.2415 0.3365 3.1255 0.3715 0.0335 2.6545 IMPDH1 0.5385 0.89950.4395 1.2965 0.331 0.5735 0.387 SLC25A4 1.5845 9.0595 1.2235 2.16251.1405 1.867 1.2185 TEGT 1.401 3.427 0.884 1.7315 0.8495 1.1535 0.6225RPML3 0.5955 0.7495 0.296 0.7785 0.289 0.637 0.462 MAP2K3 0.636 1.13850.431 0.741 0.7065 0.906 0.591 ESTs 0.62 3.725 0.2185 0.5475 0.4380.1825 1.017 AMID 1.699 2.6895 1.2005 2.6595 3.6495 5.6195 3.925 ARF4L1.382 8.4535 1.1145 1.645 1.01 1.508 0.885 GRIA2 0.768 1.3815 1.8260.724 0.3935 0.8915 0.487 ESTs 2.046 3.758 1.7975 2.428 2.9085 2.0090.7815 RPL10 2.766 8.6115 0.9505 1.3005 2.265 2.556 2.6805 PROCR 0.76953.6755 0.4425 0.6025 1.0405 0.852 0.9815 KIAA0174 1.1255 1.211 0.8811.5945 0.631 1.416 0.583 SLC1A5 3.931 3.868 5.366 2.341 3.6915 2.9472.5335 SF3A1 0.5965 0.733 0.406 0.8295 0.2405 0.635 0.4675 ESTs 0.8360.8115 0.387 0.6965 0.65 0.98 0.5765 ESTs 0.7075 1.7875 1.388 0.9741.4545 0.329 0.992 POH1 1.1285 2.8295 0.441 0.937 0.9295 1.2075 0.7855ACP5 0.8255 1.307 0.4865 0.521 1.0315 0.817 0.535 ADK 0.8285 1.2090.3015 0.435 0.993 0.797 0.53 EST 0.8655 0.179 0.1535 0.2685 0.6390.0795 0.645 FLJ13443 0.8275 0.5205 0.1785 0.266 0.8715 0.1775 0.303FLJ10976 0.9215 2.0805 0.85 1.592 1.117 0.9175 0.6435 GRP58 0.607 0.98553.1965 2.688 0.8735 0.544 1.415 Fibronectin 1 0.558 6.0425 0.8805 3.91850.3745 0.0895 3.237 PTP IVA 0.422 1.3295 0.422 1.0725 0.594 2.07751.0525 ESTs 0.7165 0.904 0.1415 0.3345 0.367 0.115 0.5965 MCT-1 1.43851.292 0.546 1.4685 1.293 1.1955 1.31 ESTs 0.985 1.779 0.5135 1.11650.5125 0.7995 0.7305 PTGS2 0.0625 0.1975 0.024 0.0715 0.021 0.0145 0.193ESTs 0.7395 1.495 0.371 0.813 0.6285 0.218 0.873 MAM 0.6575 5.7525 0.421.218 1.116 0.698 1.0755 SFRS3 1.708 3.2255 1.4135 1.2645 1.758 2.1041.325 TUBB 0.911 1.3455 0.226 0.506 1.0425 0.9855 0.9375 ESTs 0.97850.986 1.0615 1.086 0.883 1.6875 1.4255 1.225 3.9395 2.966 1.942 3.18451.92 1.091 THBD 0.111 2.3235 0.3525 0.2235 0.244 0.2945 0.1125 SLC2A10.6725 0.3475 0.198 0.531 0.7935 1.893 0.2185 CD59 0.865 2.3895 0.821.177 0.961 0.8495 1.922 ESTs 0.6335 1.144 0.088 0.2355 0.3225 0.1150.481 PRDX2 0.721 1.848 2.8795 2.5795 1.1035 2.311 0.767 ESTs 1.371515.3045 3.606 1.223 1.013 1.4025 2.688 EIF2B2 0.4645 0.9435 0.233 0.3870.359 0.49 0.4835 ESTs 1.071 3.8935 0.7155 0.9165 1.4355 0.8805 0.906EST(MTT-1B) 1.047 0.391 0.162 0.2235 0.7335 0.084 0.6565 HIP2 0.60651.079 0.5295 0.761 0.634 0.8495 0.545 TCCCIA00427 1.2655 1.59 1.5013.332 0.783 0.7435 0.5815 HARS 1.207 1.187 0.634 1.1255 0.9515 1.2731.015 HDAC3 1.1185 1.8585 0.8905 1.0185 0.6555 1.1155 0.618 ZFP92 0.8371.59 0.5205 1.409 0.559 1.411 0.572 KIAA0700 0.7185 1.694 0.5715 0.6661.2795 0.5085 0.4635 ESTs 0.8555 0.961 0.2585 0.756 0.7315 0.389 0.717NRBF-2 0.9375 2.587 0.2265 0.5405 0.959 0.526 0.683 MAX-IP1 0.72953.3305 0.597 1.407 0.577 0.762 0.366 DNMT1 1.0945 0.8405 2.0275 2.15850.8315 0.954 1.284 PRO1659 1.027 3.599 0.187 0.275 0.842 0.021 1.444PERQ1 1.0965 4.796 1.0245 3.2815 1.568 1.1255 1.252 FLJ10330 1.2551.5485 0.486 0.833 1.04 1.6045 1.177 ESTs 1.382 2.779 2.1015 2.0485 1.251.8075 1.2765 ART4 0.604 3.014 0.7945 1.3 0.393 0.6045 0.686 ACADM1.6325 1.4885 0.3485 0.8055 0.612 0.477 1.179 COX6B 1.682 6.4865 3.0652.678 1.67 1.9215 1.21 EIF4A2 1.394 6.512 0.4635 2.4105 0.792 0.94550.911 FACL3 1.403 1.8865 0.5105 1.947 2.4115 1 1.1605 RAP1B 1.142 0.79350.3005 0.6745 0.535 1.029 1.0795 PPP2R5C 0.7065 1.481 0.565 1.00151.1035 1.545 0.7925 ESTs 0.536 1.083 0.442 3.847 0.349 0.2975 0.538SNAPC5 0.965 3.2825 0.449 1.2875 0.7585 1.5055 0.3945 ARF1 1.02 4.03650.4755 1.469 0.586 1.2805 0.985 SLC20A1 1.321 3.6525 0.414 1.1745 0.2780.818 0.5445 EST(CTB2) 1.175 2.5405 2.2555 2.0525 1.285 2.805 1.4345H326 0.897 3.9665 1.95 1.2305 0.819 0.6035 0.484 FLJ20263(AKAP450) 0.7387.9685 1.9345 2.2725 0.603 0.8755 1.638 DUSP5 0.4535 0.484 0.51 1.22750.156 0.062 0.061 PFKL 1.063 3.658 0.286 0.419 1.109 0.743 0.74 JUN0.4975 1.1825 0.8815 0.365 0.6585 0.823 0.8815 SAT 0.491 0.642 0.17951.3215 3.9775 0.111 0.469 GNAS1 0.892 3.6785 0.6385 0.7105 0.6795 1.28951.128 GLUD1 1.213 0.9235 2.0105 2.4145 0.8475 0.81 1.265 TAF2F 0.96651.076 0.3775 0.575 0.443 0.998 0.5435 NUP54 1.1435 1.0595 0.253 0.59550.93 1.2285 1.196 PVALB 0.803 3034.015 29.026 34162.963 4.5445 2.88053.5565 ESTs 0.92 0.999 0.289 0.4215 0.414 0.854 0.685 EST(G3PDH) 0.9623.451 0.259 0.421 0.3485 0.8045 0.54 DKFZP434G032 0.6985 6.512 89.95722.3825 1263.422 1.834 5.4505 EIF2S2 1.0715 0.6975 0.2215 0.4095 1.340.5935 0.661 DKFZp586C1817 1.298 4.086 0.332 0.778 0.5305 0.317 1.083LOC56966 0.919 3.756 2.7275 4.3095 1.323 1.782 0.8035 CANX 1.696 0.78850.5045 1.3235 1.6535 1.408 1.1875 HNRPA1 1.5505 0.478 0.589 1.0015 0.8821.5625 1.062 FOXM1 0.854 2.216 0.4285 0.5505 0.76 0.9105 1.1685 ACTN30.838 2.366 0.6445 0.709 0.4335 0.541 0.671 EIF4E 0.6855 1.0055 0.19650.5985 0.5445 1.118 0.568 RPC62 2.002 3.1745 1.7305 1.367 4.4425 2.5061.769 STAT5B 0.8755 3.293 1.177 0.9845 0.6425 0.5175 1.269 MADD 0.87353.8565 1.0775 2.284 0.6735 1.2 0.606 TSPAN-3 0.7655 1.165 1.198 1.58350.921 0.929 0.6765 RY1 0.869 1.419 0.557 2.081 0.9705 1.567 0.6545 TTC10.9655 4.09 1.0065 0.7975 0.664 0.5235 0.673 TCEA2 1.0405 3.398 1.95752.3155 1.239 0.6775 0.8565 CD68 0.963 2.552 0.444 1.186 0.204 0.270.2555 KIAA0973 1.554 4.203 1.791 1.516 1.752 1.1825 1.133 BRE 1.02253.466 1.002 1.482 0.8155 0.6545 0.6075 CTBP1 1.0835 4.6745 3.0685 1.5711.2955 1.6935 1.3785 SMARCD2 1.07 6.044 3.235 2.036 2.4705 1.552 1.0145ACY1 1.5615 3.971 1.0215 1.689 0.9565 0.9915 0.654 BCAR1 0.7235 1.0590.587 0.633 0.801 1.277 0.56 LAPTM5 0.7885 2.657 1.379 0.8495 0.23750.2335 0.229 NDUFS1 1.082 3.4775 0.5085 1.333 1.171 1.4045 0.8045 AMFR1.5635 4.083 1.4875 3.179 1.9635 2.4385 1.5355 TMSB4X 1.2285 0.476 0.1650.2335 0.872 0.559 1.877 BCAT2 1.1685 0.12 0.1615 0.2255 1.442 0.67451.9365 PPIB 1.3855 1.404 2.69 1.053 0.6775 1.278 2.1405 CHAF1B 0.834.714 1.1595 3.514 2.2375 2.71 1.473 EFNB1 1.2285 2.168 0.9755 1.1621.6805 1.826 3.9375 OS4 1.2625 2.4505 1.0495 2.309 1.038 1.5465 1.335PSMD9 0.8785 2.708 0.539 1.236 0.73 0.8435 0.401 DHPS 1.1055 2.0531.2455 1.435 1.2985 1.235 0.924 PRSC1 0.3985 2.1255 0.341 1.0805 0.5220.499 0.9825 PXN 0.85 1.794 0.271 0.324 0.4075 0.3385 0.321 CLDN4 5.8566.512 7.0405 3211.802 56.151 30.974 1.379 FLJ10491 1.4705 6.512 0.5363.4925 1.51 1.7855 3.243 COMT 1.056 2.4605 0.505 0.905 0.709 0.69150.962 CLTH 0.73 1.967 0.4775 0.69 1.009 0.864 0.7125 TRIP7 1.0095 2.80651.123 1.442 2.3765 1.3995 0.797 KIAA0188 1.4985 1.3005 0.601 1.1950.5375 0.298 0.75 SP100 0.827 3.287 0.855 1.373 0.6015 0.5465 0.72EIF2S3 2.369 14.7625 2.0405 3.6135 4.507 3.2215 3.625 E2F4 1.193 2.00050.7935 0.955 0.729 1.0715 0.6285 GCN5L2 0.658 1.792 0.5175 0.575 0.2980.3075 0.4975 PSMD4 1.1585 5.0575 3.592 1.7545 1.3555 1.7 1.2155 HSPC1261.401 2.18 0.63 1.098 0.836 1.199 1.346 NR1D1 0.2985 4.848 0.84 3.0610.375 0.206 0.4425 RBBP2 1.479 6.402 2.1615 3.8545 1.5485 1.958 2.908PRO2605 0.901 1.724 0.658 1.3111 2.235 1.0535 0.5415 ZNF162 1.72254.0705 1.985 1.5815 1.8715 1.17 1.0205 HNRPC 1.165 1.2015 0.351 0.9490.739 0.8545 0.9 PSMD2 0.6105 1.1135 0.3255 0.6645 0.313 0.5505 0.3975PSMD2 0.552 0.7245 0.332 0.6755 0.354 0.565 0.389 HNRPD 0.707 1.35352.669 3.918 0.8065 0.8555 1.2305 MNAT1 0.637 3.608 0.3295 0.721 0.3260.6645 0.419 SCNN1A 1.745 20.372 3.2285 4.603 30.8785 1.8085 2.378 GSS0.379 1.2795 1.039 0.4315 0.1675 0.2085 0.475 DRG2 0.623 1.545 0.48750.797 0.3885 0.562 0.431 ADSL 1.4975 2.099 0.483 0.8595 0.7255 0.75450.7115 GS3955 1.0835 11.633 3.84 3.052 1.99 0.3595 0.907 DLD 0.68950.941 3.2445 3.756 0.871 1.072 1.3605 PTK7 0.453 4.8115 1.469 0.7841.893 0.6335 1.8735 PPP1R7 1.4795 4.766 2.2 3.2585 2.2935 1.287 1.175PRKCBP1 0.724 3.5355 0.701 1.0465 0.7785 1.212 1.093 SMARCA2 2.0865.0805 3.333 2.851 2.124 1.235 0.3525 MX1 1.1255 13.161 11.482 7.655511.108 1.24 38.2705 ACTR1A 1.169 4.803 0.9565 1.267 0.7945 0.91 0.8815ARPC1A 0.86 8.3555 1.001 1.348 0.7215 0.8435 0.616 NQO2 1.069 0.78250.1415 0.126 0.2615 0.055 0.049 HSJ2 1.142 2.51 0.9115 1.985 0.72551.4695 1.8275 IFI16 0.5615 1.5035 0.275 0.204 0.019 0.016 0.3175 TOP2A0.6605 0.3715 0.181 0.4205 0.82 0.844 0.5655 HMGCL 0.924 3.8995 1.14751.1965 0.8875 0.858 0.721 MGAT2 1.5065 1.1425 0.547 0.6215 0.3465 0.6110.993 PABPC1 0.6095 2.278 0.5095 0.3195 0.469 0.2935 0.7905 MNPEP 1.24150.7725 1.6045 1.8895 0.7735 1.037 1.3945 P130 1.043 1.3645 0.2815 0.6810.625 0.6075 1.689 DUSP12 1.1955 1.7505 1.0515 0.9915 2.0275 1.01550.7885 UQCRC2 1.099 2.431 0.5465 1.148 0.94 2.072 0.858 SLC25A6 1.71212.1205 1.356 2.082 1.161 2.0135 1.3713 LGALS3 0.739 5.6375 0.461 0.15550.602 0.151 1.371 FSFT1 0.8085 0.697 0.876 0.704 0.749 0.935 1.0365TMSB4X 1.238 0.1725 0.19 0.244 1.331 0.6875 2.1755 DCTN1 0.9165 3.34750.7175 1.7875 0.6715 0.99 0.573 DXS1357E 1.2195 4.5985 1.069 1.69450.933 1.2605 1.012 RPL18 2.145 6.2765 2.634 1.9115 1.002 1.9055 1.202HBG1 1.408 1.973 0.463 0.779 0.976 1.5835 0.947 CTNNA1 1.023 0.82950.295 0.57 0.1075 0.5995 0.521 PGAM1 1.035 1.873 0.657 0.7745 0.30950.6495 0.927 PRO2706 1.066 1.6325 0.5065 1.412 0.9995 1.1125 0.818 CCT41.0075 0.2765 0.0505 0.4785 0.8575 1.2675 0.393 KIAA0106 1.243 0.67150.3915 0.845 1.1175 0.754 0.6045 UBE1 0.848 3.496 1.2705 3.717 2.16552.7045 1.523 ZNF220 1.2655 2.1565 1.451 6.1565 0.855 0.9455 1.333 SOD11.279 2.449 1.3525 2.821 2.0195 1.3995 0.7495 PFKP 1.3625 4.3135 0.7630.7515 1.092 1.319 0.9995 SHMT2 2.79 2.6455 0.6015 0.4655 1.561 1.21050.9965 GPI 1.16 1.4485 1.2495 1.3795 1.229 1.0365 1.1715 CALM3 1.03653.228 0.779 1.0685 0.8625 0.8105 0.733 RPS8 1.876 3.672 0.974 1.8761.696 1.4415 1.407 PTS 1.25 1.9595 1.036 1.4485 2.795 0.9325 0.9975HNRPL 0.963 1.693 1.4435 1.281 1.511 1.8605 1.2505 CKB 0.252 3.54251.5175 0.653 0.1605 1.278 0.2115 HADHA 1.1375 3.8725 0.326 0.424 1.1850.687 0.747 RPS27 1.189 5.5065 1.9225 1.4805 1.096 1.156 0.771 CCT6A1.4345 1.3275 0.6925 1.0815 3.487 1.0275 0.9125 Gene Name MB134 MB157MB436 MB453 BT20 BT474 BCTis-3 TNC 0.083 0.229 0.2915 1.322 0.0275 0.310.862 ALDOC 0.6725 0.084 0.477 0.846 0.5965 0.3565 1.0645 ALCAM 1.25350.193 0.299 2.2875 0.0715 0.607 0.248 NCBP2 0.823 0.3445 0.394 0.31650.6855 0.709 0.9885 LOC57862 1.4175 1.389 0.753 1.8475 1.212 1.746 0.953karyopherin a2 0.536 1.156 0.672 1.1825 0.913 1.327 0.9935 ESTs 5.37453.409 0.5135 2.5625 1.2645 2.3425 1.269 FLJ10509 1.4435 0.6365 0.91251.3555 1.6975 1.211 1.487 PRPSAP1 2.1275 2.588 0.828 2.205 1.626 1.4873.9915 Fibronectin 1 0.052 7.429 0.6615 0.047 0.2645 0.0825 14.9675IMPDH1 0.746 0.294 0.3385 0.3705 0.7115 0.739 0.788 SLC25A4 1.15553.7145 1.3925 1.3215 0.5115 2.456 1.3635 TEGT 2.5555 0.7165 0.39451.3355 1.243 1.5195 1.5285 RPML3 0.75 0.2245 0.3375 0.3575 0.733 0.74450.7515 MAP2K3 0.8695 1.1805 0.8895 0.533 1.0785 0.7475 0.6815 ESTs 0.2050.711 0.376 0.1465 0.1745 0.143 0.437 AMID 4.7245 4.1005 6.505 2.04256.565 3.195 1.4385 ARF4L 0.9695 2.686 1.1445 1.018 0.604 1.83 1.2565GRIA2 1.084 0.488 0.365 1.245 0.7055 0.7945 0.8765 ESTs 1.709 1.34451.0245 1.998 2.459 3.2003 1.6245 RPL10 1.3545 1.761 3.2775 5.325 1.1721.668 2.157 PROCR 0.6095 1.3195 0.9655 0.6315 1.338 0.534 0.9085KIAA0174 1.2295 0.7765 0.6725 0.502 2.6715 1.483 0.993 SLC1A5 6.25752.794 2.0725 3.821 2.165 2.8855 1.453 SF3A1 0.7375 0.293 0.3325 0.2980.716 0.6905 0.714 ESTs 0.53 0.0445 0.563 0.291 0.677 0.671 0.8965 ESTs0.98 1.7145 1.764 0.3215 0.346 0.8265 2.1375 POH1 1.282 0.707 1.10350.821 1.447 1.0855 0.924 ACP5 0.5365 0.4775 1.424 0.4095 0.7035 0.8720.5625 ADK 0.639 0.4465 0.3545 0.424 0.7015 0.992 0.4065 EST 0.3 0.1470.2525 0.2465 0.165 0.11 0.122 FLJ13443 0.224 0.584 0.1705 0.653 0.20.098 0.344 FLJ10976 1.1 0.851 0.705 0.921 2.1265 1.29 2.2155 GRP582.3025 1.113 1.101 2.3115 1.8155 0.762 1.4895 Fibronectin 1 0.211 10.6240.691 0.114 0.2675 0.076 32.204 PTP IVA 1.931 1.3455 0.6125 0.88250.2185 1.107 1.0365 ESTs 0.539 0.7465 0.4645 0.1635 0.2185 0.1845 0.817MCT-1 1.5805 5.669 1.3235 0.516 1.963 1.1625 0.958 ESTs 0.758 0.960.8875 0.584 0.456 0.523 0.8465 PTGS2 0.044 0.0095 0.0815 0.0495 0.01250.028 0.013 ESTs 0.359 1.08 0.7935 0.175 1.193 0.5335 1.8195 MAM 0.73054.4885 0.443 0.3025 0.23 0.5795 2.241 SFRS3 1.511 1.7895 1.3305 1.39751.632 2.5915 2.3715 TUBB 0.4865 0.578 0.4805 0.3135 0.5895 0.867 0.935ESTs 1.476 0.8335 0.935 1.194 1.0705 3.561 2.8205 1.843 1.4565 0.5541.357 2.8505 5.027 1.082 THBD 0.1645 0.0075 0.151 0.3495 0.0405 0.0890.1725 SLC2A1 3.522 0.817 0.502 0.344 0.5585 0.8135 0.855 CD59 0.79252.476 0.7755 0.734 0.735 0.8725 7.412 ESTs 0.114 1.0425 0.3505 0.09550.1055 0.158 0.7315 PRDX2 0.501 0.1875 1.208 1.525 0.8105 2.3125 0.787ESTs 3.729 0.3425 1.6685 0.993 4.8455 5.5175 1.9915 EIF2B2 0.5015 1.210.295 0.364 0.2235 0.435 0.328 ESTs 0.9505 1.981 1.319 0.246 1.01 0.7640.9785 EST(MTT-1B) 0.2805 0.296 0.3035 0.197 0.2125 0.1045 0.1745 HIP20.8555 1.167 0.8825 0.506 0.9325 0.7075 0.6435 TCCCIA00427 1.7045 1.33551.066 1.752 1.272 2.3445 1.254 HARS 0.834 1.173 1.0685 1.2255 1.14751.0605 1.3 HDAC3 1.1245 1.085 0.8495 0.571 0.898 1.044 0.7 ZFP92 0.94151.1055 0.603 1.1 0.6825 5.3125 2.365 KIAA0700 0.512 1.3745 0.7875 0.38551.117 0.7535 0.457 ESTs 0.9295 1.2825 0.733 0.403 0.5775 0.5245 0.4285NRBF-2 0.7695 0.819 0.95 0.6775 0.644 0.599 0.53 MAX-IP1 1.0845 0.84250.397 0.4715 0.4095 0.3235 1.5375 DNMT1 1.7225 0.8565 1.044 2.06051.5975 0.754 1.1775 PRO1659 0.099 0.921 0.0595 0.5335 0.0485 0.07 1.159PERQ1 1.5895 5.2495 1.145 1.556 1.03 3.25 2.3 FLJ10330 0.7975 1.8080.559 1.5015 1.8595 1.081 1.4795 ESTs 1.3085 1.2165 1.022 0.744 0.91651.873 1.553 ART4 1.066 0.4065 0.518 0.391 0.8325 0.902 1.1305 ACADM2.2965 0.821 2.154 0.4005 1.266 0.8335 2.1475 COX6B 2.76 5.2755 1.41651.1105 2.1125 3.3935 2.335 EIF4A2 0.64 0.002 0.3925 2.472 1.831 1.0811.3555 FACL3 4.1225 3.1055 0.893 3.682 1.4815 3.4015 1.081 RAP1B 1.03650.9895 1.3155 0.5945 0.9845 0.7125 1.6955 PPP2R5C 0.949 1.0355 0.79450.906 1.0525 1.3081 1.4955 ESTs 0.7285 0.393 0.254 0.279 0.3775 0.52450.3985 SNAPC5 1.091 0.945 1.3685 0.5625 1.384 1.0605 0.662 ARF1 0.51350.194 0.804 1.633 1.467 0.4765 0.8595 SLC20A1 0.533 0.0015 0.851 0.53550.3375 0.2725 0.489 EST(CTB2) 3.8435 2.0365 0.8045 2.281 0.934 1.8013.129 H326 1.026 0.7975 0.616 0.5505 0.666 1.2165 1.6595FLJ20263(AKAP450) 2.2365 2.7495 0.735 0.67 0.8525 1.4945 2.4845 DUSP50.218 0.096 0.1545 0.2435 0.037 0.0425 0.304 PFKL 0.955 1.388 0.8590.8835 0.918 0.4175 0.447 JUN 0.1985 0.767 0.773 1.0665 0.507 0.68351.2975 SAT 0.6935 0.1755 0.3135 0.2115 0.225 0.627 1.149 GNAS1 0.81.0245 0.9545 1.253 2.394 1.083 0.9915 GLUD1 1.7325 0.777 0.947 2.331.265 0.795 1.0165 TAF2F 0.602 1.6335 0.4445 0.539 0.8575 0.8705 1.049NUP54 0.8125 0.826 0.9855 1.041 1.152 1.7865 0.837 PVALB 5.18 9.35752.5555 2.3745 2.8045 6.814 4.736 ESTs 0.4155 0.1175 0.7225 0.4055 1.02550.6755 0.5645 EST(G3PDH) 0.31 0.117 0.657 0.3365 0.9385 0.5835 0.5265DKFZP434G032 4.919 9.817 1.4585 2.224 2.4305 2.6 10279.539 EIF2S2 0.92751.254 0.7515 0.7625 0.847 0.293 0.373 DKFZp586C1817 1.0435 0.683 0.42450.145 1.73 0.443 1.008 LOC56966 2.044 2.3915 0.757 0.7455 2.525 2.7411.254 CANX 2.065 2.773 2.262 1.528 1.2645 1.365 0.4385 HNRPA1 0.6970.5545 0.882 1.1935 0.788 0.8645 0.956 FOXM1 0.4075 0.2225 1.927 0.5061.124 0.949 1.111 ACTN3 0.7415 0.4775 0.5715 0.619 0.3935 0.5585 0.9855EIF4E 0.7465 0.5455 0.701 1.369 0.937 0.9685 0.553 RPC62 1.2155 1.89252.1285 0.976 2.1585 1.261 3.415 STAT5B 0.9355 1.9855 0.6505 0.42150.5355 0.756 0.6965 MADD 0.9985 0.829 0.4595 0.606 0.6385 0.781 1.221TSPAN-3 2.062 1.3325 0.468 0.0535 0.278 0.1785 0.7485 RY1 1.142 1.32751.419 0.9495 0.936 0.8085 0.9415 TTC1 1.0205 1.882 0.7765 0.6485 0.49450.7855 0.6875 TCEA2 0.9595 2.5575 0.375 0.5555 1.7475 1.577 0.7495 CD680.246 1.02 0.4565 0.19 0.325 0.248 1.716 KIAA0973 2.847 1.851 1.411.1345 1.404 3.31 1.978 BRE 1.341 1.467 0.736 0.62 0.737 0.878 1.261CTBP1 1.6185 1.4985 2.015 1.4565 1.1645 1.4155 2.496 SMARCD2 2.0051.2945 0.28 1.42 2.3675 4.6585 0.3875 ACY1 0.622 0.526 0.4715 0.8551.084 1.0755 0.925 BCAR1 1.102 1.231 0.6615 1.2765 0.6835 0.7295 0.7375LAPTM5 0.459 0.1235 0.2265 0.3665 0.1855 0.146 1.895 NDUFS1 1.29351.4475 1.171 0.83 1.1985 1.193 1.0725 AMFR 2.039 1.5275 1.252 1.9932.527 1.3955 1.577 TMSB4X 0.359 1.0855 0.86 0.1465 0.1915 1.476 1.1765BCAT2 0.467 2.32 0.816 0.1215 0.168 1.5805 1.7565 PPIB 2.4055 2.7531.6725 2.5865 1.3085 0.533 1.289 CHAF1B 2.8405 2.8915 1.8245 1.43851.4295 2.78 2.123 EFNB1 2.488 3.235 1.4295 0.8675 0.6145 1.9215 2.2775OS4 1.8455 0.979 0.5125 2.106 1.167 1.5885 3.1285 PSMD9 0.905 0.83050.851 0.5325 0.5155 0.9055 0.988 DHPS 0.6635 1.165 0.923 0.868 0.7611.175 0.6 PRSC1 0.9015 1.2235 0.287 0.308 0.673 0.331 2.384 PXN 0.5430.39 0.6675 0.39 0.678 0.529 0.5175 CLDN4 5.72155 5.215 1.543 9.81728.0275 136.3065 4175.7835 FLJ10491 1.4965 0.006 2.1085 0.45 2.6342.1025 1.371 COMT 0.5755 0.7285 0.6055 0.683 0.7345 0.777 0.916 CLTH0.495 0.7515 0.9225 0.669 1.188 0.5015 0.905 TRIP7 2.5205 1.942 1.82651.3735 0.5195 2.927 1.4265 KIAA0188 0.9945 2.7635 0.596 0.904 0.60050.736 1.779 SP100 1.148 2.4075 0.502 0.3085 0.6435 0.291 1.857 EIF2S33.7685 4.4755 4.35 2.4815 3.083 5.41 5.58 E2F4 1.0375 0.4285 0.74350.7525 1.846 1.1245 1.037 GCN5L2 0.627 2.0175 3.049 0.206 0.214 0.2880.9845 PSMD4 1.5415 1.667 2.1205 1.345 1.2155 1.497 2.6205 HSPC126 1.4630.493 1.4065 1.563 1.875 1.4075 1.557 NR1D1 0.613 0.983 0.45 0.2430.2725 0.1655 2.318 RBBP2 2.299 1.3615 4.235 2.2035 3.2825 2.618 3.394PRO2605 15.176 1.2685 0.4715 0.6135 0.5905 1.4295 0.666 ZNF162 1.95252.015 1.765 1.14 1.2475 3.7065 2.343 HNRPC 0.7345 0.553 1.094 1.2470.9495 0.879 0.8185 PSMD2 0.7405 0.2575 0.361 0.3135 0.6915 0.6895 0.758PSMD2 0.7315 0.2665 0.311 0.3305 0.704 0.6935 0.649 HNRPD 2.118 1.06251.042 2.2275 1.763 0.8475 1.3195 MNAT1 0.7715 0.2935 0.3705 0.417 0.7840.7525 0.7295 SCNN1A 71.293 2.253 2.0385 1.5535 30.312 19.2445 4.658 GSS0.205 0.2935 0.3145 0.3595 0.269 0.203 0.3285 DRG2 0.7805 0.2655 0.4210.3535 0.6865 0.72 1.04 ADSL 0.9785 1.2665 0.7895 0.6425 0.553 0.9880.946 GS3955 1.6365 1.6395 1.1165 0.62 1.1645 0.356 1.472 DLD 2.2291.1315 1.05 2.6815 1.805 0.965 1.273 PTK7 0.8635 1.899 0.368 0.542 2.021.1355 3.0105 PPP1R7 2.95 2.09 1.1385 1.368 1.981 1.947 2.1225 PRKCBP11.304 1.3 0.713 1.4 1.7865 1.563 1.526 SMARCA2 3.287 1.0865 1.51451.4175 2.0375 0.5805 3.353 MX1 9.4925 177.459 5.1435 3.2365 6.85352.7345 17.302 ACTR1A 0.602 0.662 1.055 0.561 0.762 0.776 0.908 ARPC1A1.1655 0.5525 0.712 0.771 1.0795 0.9775 1.057 NQO2 0.073 0.0275 0.1450.064 0.139 0.031 0.6335 HSJ2 1.268 1.8435 1.872 1.81 0.719 1.747 2.3705IFI16 0.082 2.3755 0.631 0.0455 0.0685 0.0295 1.175 TOP2A 0.2155 0.14450.4195 0.1865 0.61 0.6195 0.769 HMGCL 0.648 1.1385 1.1415 0.5735 0.8390.9715 1.1035 MGAT2 0.5085 0.629 0.3385 1.029 0.5015 0.438 0.532 PABPC10.4085 0.957 2.0415 0.3885 0.5955 0.33 0.69 MNPEP 1.697 0.9545 0.92451.6125 1.3275 0.757 1.0515 P130 0.4415 0.716 0.8015 0.451 0.267 0.6671.3695 DUSP12 1.057 1.1505 1.004 0.92 1.0685 1.9875 1.592 UQCRC2 3.00751.551 1.834 1.178 1.097 1.759 1.0965 SLC25A6 1.2885 4.3495 1.6095 1.40650.472 2.906 1.522 LGALS3 1.6405 2.215 0.5115 0.27 0.75 0.8815 1.233FSFT1 1.2785 0.9515 0.608 1.719 0.767 0.6835 0.7845 TMSB4X 0.514 2.5230.8575 0.1475 0.1925 1.693 1.629 DCTN1 1.0465 0.9455 0.494 0.467 1.03550.443 1.3535 DXS1357E 0.8015 1.354 1.111 1.005 0.8185 1.3365 0.9385RPL18 0.91 0.6875 0.914 0.859 1.5585 1.5095 1.271 HBG1 0.4 0.8015 0.99450.7335 0.645 0.8825 0.615 CTNNA1 1.4065 0.0005 0.309 0.6765 0.795 1.21650.7585 PGAM1 0.5585 0.507 0.4985 0.414 0.328 0.587 0.8925 PRO2706 2.1721.4275 0.7875 1.065 1.4985 1.67 1.056 CCT4 0.4345 0.608 0.9985 0.45551.1 0.549 0.8025 KIAA0106 0.8625 0.7125 0.8905 0.51 1.583 1.272 0.868UBE1 2.7625 2.7385 1.754 1.391 1.324 2.75 2.168 ZNF220 1.349 1.57350.6875 1.0335 0.969 1.2055 2.333 SOD1 1.633 2.692 2.051 1.0645 2.52253.224 2.2375 PFKP 1.37 1.374 0.6555 0.7935 1.5745 1.422 1.168 SHMT21.106 1.502 0.717 1.0935 0.6265 0.969 1.5105 GPI 1.981 0.804 0.368 1.1261.1215 1.0235 0.988 CALM3 0.5515 0.9525 0.8255 0.621 0.7405 0.889 1.0095RPS8 0.9335 2.07 2.18 1.119 0.867 1.7025 1.418 PTS 1.498 1.5405 1.38650.893 1.5505 1.363 3.1175 HNRPL 1.26 1.499 0.771 0.9405 1.6075 2.3 1.939CKB 1.374 0.8095 0.1075 2.6755 0.111 0.0265 1.051 HADHA 1.1565 1.48650.8275 0.7885 0.8945 0.358 0.615 RPS27 0.585 1.8435 1.2445 1.0555 0.67951.972 2 CCT6A 1.2785 2.0385 1.201 1.0285 3.4195 2.9105 1.096

[0048] The high quality cDNA microarrays were then used to measureexpression of 768 arrayed elements in malignant breast cancer cell lines(n=10) and tissue samples (n=3) using the non-tumorigenic cell lineMDA/H6 as a common reference. The name and origin of the breast cancercell lines and tissues are listed in Table 3. Pearson coefficient ofcorrelation was used to compute the matrix of similarities anddissimilarities between samples and genes. The complex matrixrelationships between 15 cancer samples and between 202 genes weresimplified and visualized by multidimensional scaling and hierarchicaldendrogram clustering analyses. First, the expression profiles of 202genes in two MDA-MB-231 samples were essentially identical (r=0.982) andthe expression pattern of the melanoma sample was the most dissimilar tothat of MDA-MB-231 (r=0.325), as expected. Secondly, the expressionprofiles of other 12 breast cancers were distributed between theidentity and the dissimilarity and had their own expression patterns,demonstrating the extensive heterogeneous nature of these breast cancercells. Thirdly, the expression profiles of BT20, BT474 and ZR75-1 celllines were more similar to each other (r=0.796) than to others. TABLE 3Human Cancer Cell Line and Tissue Original Name Symbol ClinicalDiagnosis Source MDA-MB-231 MB231 Adenocarcinoma ATCC¹ MDA/H6 MDA/H6Non-tumorigenic Dr. Negrini³ MDA-MB-134 MB134 Carcinoma ATCC MDA-MB-157MB157 Carcinoma ATCC MDA-MB-436 MB436 Adenocarcinoma ATCC MDA-MB-453MB453 Carcinoma ATCC MDA-MB-468 MB468 Adenocarcinoma ATCC BT-20 BT20Adenocarcinoma ATCC BT-474 BT474 Ductal carcinoma ATCC BT-549 BT549Ductal carcinoma ATCC ZR75-1 ZR751 Ductal carcinoma ATCC Breast cancertissue 1 BCTis-1 Poorly differentiated LCC² invasive ductal carcinomaBreast cancer tissue 2 BCTis-2 Poorly differentiated LCC infiltratingductal carcinoma Breast cancer tissue 3 BCTis-3 Poorly differentiatedLCC infiltrating carcinoma Melanoma tissue MelTis Metastatic malignantLCC melanoma

[0049] The microarray gene expression analysis revealed 19 genes withhigh frequent alterations in their expression in human breast cancers.Out of the 19 genes, 9 genes were over-expressed (Table 4) and 10 geneswere under-expressed (Table 5) in breast cancers at the frequenciesgreater than 77% (n=13). TABLE 4 Over-expressed BCSGs in breast cancercell line and tissue samples Symbol Locus ID Nucleotide Seq. Amino acidSeq. MX1 4599 SEQ ID NO:1 SEQ ID NO:20 PVALB 5816 SEQ ID NO:2 SEQ IDNO:21 RBBP2 5927 SEQ ID NO:3 SEQ ID NO:22 AIF 84883 SEQ ID NO:4 SEQ IDNO:23 CLDN4 1364 SEQ ID NO:5 SEQ ID NO:24 KRT23 25984 SEQ ID NO:6 SEQ IDNO:25 SLC1A5 6510 SEQ ID NO:7 SEQ ID NO:26 EIF2S3 1968 SEQ ID NO:8 SEQID NO:27 SCNN1A 6337 SEQ ID NO:9 SEQ ID NO:28

[0050] TABLE 5 Under-expressed BCSGs in breast cancer cell line andtissue samples Symbol Locus ID Nucleotide Seq. Amino acid Seq. THBD 7056SEQ ID NO:10 SEQ ID NO:29 PTGS2 5743 SEQ ID NO:11 SEQ ID NO:30 GSS 2937SEQ ID NO:12 SEQ ID NO:31 DUSP5 1847 SEQ ID NO:13 SEQ ID NO:32 NQO2 4835SEQ ID NO:14 SEQ ID NO:33 TNC 3371 SEQ ID NO:15 SEQ ID NO:34 LAPTM5 7805SEQ ID NO:16 SEQ ID NO:35 IFI16 3428 SEQ ID NO:17 SEQ ID NO:36 CD68 968SEQ ID NO:18 SEQ ID NO:37 EIF2B2 8892 SEQ ID NO:19 SEQ ID NO:38

[0051] Six of the nine over-expressed genes were known to be involved inhuman cancers. The interferon-inducible protein p78 (MX1) isover-expressed in human prostate cancer cell line LNCaP (Vaarala et al.,Lab. Invest., 80:1259-1268, 2000). Parvalbumin (PVALB) is a Ca²⁺ bindingprotein and was highly expressed in human carcinoma, mouse neuroblastomaand rat glioma (Pfyffer et al., 412:135-144, 1987). The retinoblastomabinding protein 2 (RBBP2) can bind to the tumor suppressor gene RB andreverse RB-mediated suppression of the activity of the E2F transcriptionfactor (Kim et al., Mol. Cell Biol., 14:7256-7264, 1994). The apoptosisinducible factor (AMID) is a flavoprotein that is normally confined tomitochondria and is sufficient to induce apoptosis of isolated nuclei(Susin et al., Nature, 397:441-446, 1999). Claudin 4 (CLDN4) is a memberof the family of tight junction proteins and was shown to up-regulatedin ovarian cancer (Hough et al., Cancer Res., 60:6281-6287, 2000). Anexpression of keratin 23 (KRT23) was highly inducible by pro-apoptoticagent sodium butyrate in different pancreatic cancer cells and thisinduction was blocked by expression of p21 (WAF1/CIP1) antisense RNA(Zhang et al., 30:123-135, 2001). In addition, soluble carrier family 1member 5 (SLC1A5) is a neutral amino acid transport-like protein and wasup-regulated in 12 of the 13 breast cancer cell lines/tissue samples.Eukaryotic translation initiation factor 2B gamma (EIF2S3) and sodiumchannel nonvoltage-gated 1α (SCNN1A) were up-regulated in 12 and 10 ofthe 13 breast cancer cell lines/tissue samples, respectively.

[0052] Among the under-expressed genes listed in Table 5, thrombomodulin(THBD), a negative regulator of coagulation, was reported to involve invascular diseases and cancers. (Kim et al., Anticancer Res.,17:2319-2323, 1997; Hosaka et al., Cancer Lett., 161:231-240,2000).Prostaglandin-endoperoxide synthase 2 (PTGS2) was reported to beundetectable in mammary invasive carcinomas and was more likely detectedin ductal carcinomas in situ (Soslow et al., Cancer, 89:2637-2645,2000). PTGS2 was down-regulated in all 13 breast cancer celllines/tissue samples. Up-regulation of glutathione synthetase (GSS) isknown to increase the cellular levels of glutathione that in turnfacilitates growth of certain cells (Huang et al., FASEB J, 15:19-21,2001). The GSS expression was decreased in 12 of 13 breast cancer celllines/tissue samples, suggesting that the advanced cancer cells do notrequire high levels of glutathione for their growth. Dual specificityprotein tyrosine phosphatase 5 (DUSP5) is inducible by serum stimulationof fibroblasts and by heat shock, and the DUSP5 induction may lead tothe deactivation of mitogen- or stress-activated protein kinase 3(MAPK3) that participates in cell cycle progression (Ishibashi et al.,J. Biol. Chem., 269:29897-29902, 1994). NAD(P)H menadione oxidoreductase2 (NQO2) is expressed in human heart, brain, lung, liver, and skeletalmuscle but is not expressed in placenta, implying its decrease in fastgrowth tissue. The expression of NQO2 is inducible by antioxidants andits role in cancer remains unknown. Interestingly, the expression of theeukaryotic translation initiation factor 2 beta subunit (EIF2B2) wasdecreased more than 2 fold in 11 of 13 breast cancer cell lines/tissuesamples, whereas the gamma subunit (EIF2S3) was up-regulated in all 13breast cancer cell lines/tissue samples. The discovery that the inverselevels of EIF2B2 and EIF2S3 were associated with breast cancerprogression has not been reported before. Hexabrachion (TNC) is anextracellular matrix glycoprotein that modulates cellular organization(Talts et al. J. Cell Sci., 112:1855-1864, 1999) and the TNC expressionwas down-regulated in 10 of the 13 breast cancer cell lines/tissuesamples.

[0053] Further analysis demonstrated that the THBD RNA levels decreasedin all 13 breast cancer cell lines/tissue samples (FIG. 4, panel (B)).In addition, Western blot analysis correlated the THBD proteinexpression to its RNA levels in all five cell lines tested. Furthermore,the THBD protein levels were negative in all 18 cases of the advancedbreast cancer cells in contrast to normal mammary epithelial cells,measured by in situ immunohistochemical staining (Table 6). It thusappears that THBD expression is inversely correlated to the developmentof breast cancer. TABLE 6 Results of in situ immunohistochemicalstaining of THBD antibody on 20 cases of breast normal and cancerspecimens Pathological diagnosis THBD staining Case MalignancyMetastasis to RLN MEC BCC 1 Infiltrating ductal To 19 of 20 RLN + + + −carcinoma 2 Infiltrating ductal To 2 of 2 RLN + + + − carcinoma 3Infiltrating lobular and  To 5 of 15 RLN + + − ductal carcinoma 4Infiltrating ductal To 1 of 9 RLN + + + − carcinoma 5 Infiltratingductal  To 1 of 12 RLN + + − carcinoma 6 Infiltrating ductal NE + + + −carcinoma 7 Infiltrating carcinoma, NE + + + − poorly differentiated 8Infiltrating ductal NE + + + − carcinoma 9 Infiltrating ductal NE + + −carcinoma 10 Infiltrating ductal NE + + + − carcinoma 11 Infiltratingductal NE + + − carcinoma 12 Infiltrating ductal NE + + − carcinoma 13Infiltrating ductal NE + + + − carcinoma 14 Infiltrating ductal NE + + +− carcinoma 15 Infiltrating ductal NE + + + − carcinoma 16 Infiltratingductal NE + + − carcinoma 17 Infiltrating ductal NE + + + − carcinoma 18Infiltrating ductal NE + + − carcinoma 19 Infiltrating NE + + + + +adenocarcinoma, moderately well differentiated 20 Infiltrating ductalNE + + + + carcinoma with intramammary lymphatic invasion

[0054] Each case of the normal and breast cancer specimens was from thesame patients. All the sections were purchased from Lombardi CancerCenter (LCC) Histology Facility. The malignant diagnosis was derivedfrom LCC pathological reports and further verified using HE stainedsections. The Metastatic diagnosis was from the LCC pathologicalreports. The criteria for scoring the positive and negative results arefollows: the intensities of immunohistochemical staining: 0 (none), 1(weak), 2 (moderate), and 3 (strong); distribution of the intensities: 0(none), 1 (1-15%), 2 (26-50%), 3 (51-75%), and 4 (76-100%); sum=anintensity number+distribution number, sum 0 is score 0, sums 1, 2, and 3were grouped as score 1, sums 4 and 5 were grouped as score 2, sums 6and 7 were grouped to score 3; negative (−): means score 0 or score 1,positive (++) means score 2 and positive (+++) means score 3. THBD:thrombomodulin. MEC: normal mammary epithelial cells; BCC: breast cancercells; RLN: regional lymph nodes. NE: non evidence.

[0055] BCSG Products as Markers for Breast Cancer

[0056] Most of the BCSGs listed in Tables 4 and 5 have not beenpreviously associated with breast cancer. BCSG homologs from otherorganisms may also be useful in the use of animal models for the studyof breast cancer and for drug evaluation. BCSG homologs from otherorganisms may be obtained using the techniques outlined below.

[0057] In one aspect, the present invention is based on theidentification of a number of genes, designated breast-cancer specificgenes (BCSGs) set forth in Tables 4 and 5, which are differentiallyexpressed between the breast cancer tissues/cell lines and thenon-tumorigenic control tissues/cell lines. The proteins encoded bythese genes may in turn be components of disease pathways and thus mayserve as markers of breast cancer development or as novel therapeutictargets for treatment and prevention of breast cancer.

[0058] Accordingly, the present invention pertains to the use ofpolynucleotides transcribed from and polypeptides encoded by the BCSGsof Table 4 and 5 as markers for breast cancer. Moreover, the use ofexpression profiles of these genes can indicate the presence of or arisk of breast cancer. These markers are further useful to correlatedifferences in levels of expression with a poor or favorable prognosisof breast cancer. For example, panels of the BCSGs can be convenientlyarrayed on solid supports for use in kits. The BCSGs can be used toassess the efficacy of a treatment or therapy of breast cancer, or as atarget for a treatment or therapeutic agent. The BCSGs can also be usedto generate gene therapy vectors that inhibit breast cancer.

[0059] Therefore, without limitation as to mechanism, the invention isbased in part on the principle that modulation of the expression of theBCSGs of the invention may ameliorate breast cancer, when they areexpressed at levels similar or substantially similar to normal(non-diseased) tissue.

[0060] As an example, the expression of THBD, one of the BCSGs listed inTable 5, is dowregulated in the parental metastatic breast cancer cellline MDA-MB-231 comparing to the non-tumorigenic derivative MDA/H6.Accordingly, modulation of the down-regulated THBD gene to normal levels(e.g., levels similar or substantially similar to tissue substantiallyfree of breast cancer) may allow for amelioration of breast cancer.

[0061] In another embodiment of the invention, a BCSG product (includingpolynucleotides transcribed from a BCSG and polypeptide translated fromsuch polynucleotides) can be used as a therapeutic compound of theinvention. In yet other embodiments, a modulator of an BCSG product ofthe invention may be used as a therapeutic compound of the invention, ormay be used in combination with one or more other therapeuticcompositions of the invention. Formulation of such compounds intopharmaceutical compositions is described in subsections below.

[0062] In another aspect of the invention, the levels of BCSMs aredetermined in a particular subject sample for which either diagnosis orprognosis information is desired. The level of a number of BCSMssimultaneously provides an expression profile, which is essentially a“fingerprint” of the presence or activity of a BCSG or plurality ofBCSGs that is unique to the state of the cell. In certain embodiments,comparison of relative levels of expression is indicative of theseverity of breast cancer, and as such permits for diagnostic andprognostic analysis. Moreover, by comparing relative expression profilesof BCSGs from tissue samples taken at different points in time, e.g.,pre- and post-therapy and/or at different time points within a course oftherapy, information regarding which genes are important in each ofthese stages is obtained. The identification of genes that areabnormally expressed in breast cancer tissue versus normal tissue, aswell as differentially expressed genes during breast cancer development,allows the use of this invention in a number of ways. For example,comparison of expression profiles of BCSGs at different stages of thedisease progression provides a method for long-term prognosing,including survival.

[0063] The discovery of the differential gene expression patterns forindividual or panels of BCSMs allows for screening of test compoundswith the goal of modulating a particular expression pattern. Forexample, screening can be done for compounds that will convert anexpression profile for a poor prognosis to one for a better prognosis.In certain embodiments, this may be done by making biochips comprisingsets of BCSMs, which can then be used in these screens. These methodscan also be done on the protein level. For example, protein expressionlevels of the BCSGs can be evaluated for diagnostic and prognosticpurposes or to screen test compounds. Furthermore, the modulation of theactivity or expression of a BCSM may be correlated with the diagnosis orprognosis of breast cancer.

[0064] BCSG-Related Polynucleotides

[0065] BCSG-related polynucleotides can be prepared using any of avariety of techniques. For example, a polynucleotide may be identified,as described in more detail below, by screening a microarray of cDNAsfor tumor-associated expression (i.e., expression that is at least twofold greater in a breast tumor than in normal tissue, as described inthe present invention. Alternatively, polynucleotides may be amplifiedfrom cDNA prepared from cells expressing the proteins described herein,such as breast cancer cells. Such polynucleotides may be amplified viapolymerase chain reaction (PCR). For this approach, sequence-specificprimers may be designed based on the sequences provided herein, and maybe purchased or synthesized.

[0066] An amplified portion may be used to isolate a full length genefrom a suitable library (e.g., a breast cancer cDNA library) using wellknown techniques. Within such techniques, a library (cDNA or genomic) isscreened using one or more polynucleotide probes or primers suitable foramplification. Preferably, a library is size-selected to include largermolecules. Random primed libraries may also be preferred for identifying5′ and upstream regions of genes. Genomic libraries are preferred forobtaining introns and extending 5′ sequences.

[0067] Alternatively, there are numerous amplification techniques forobtaining a full length coding sequence from a partial cDNA sequence.Within such techniques, amplification is generally performed via PCR.Any of a variety of commercially available kits may be used to performthe amplification step. Primers may be designed using, for example,software well known in the art. Primers are preferably 22-30 nucleotidesin length, have a GC content of at least 50% and anneal to the targetsequence at temperatures of about 68° C. to 72° C. The amplified regionmay be sequenced as described above, and overlapping sequences assembledinto a contiguous sequence.

[0068] One such amplification technique is inverse PCR, which usesrestriction enzymes to generate a fragment in the known region of thegene. The fragment is then circularized by intramolecular ligation andused as a template for PCR with divergent primers derived from the knownregion. Another such technique is known as “rapid amplification of cDNAends” or RACE. This technique involves the use of an internal primer andan external primer, which hybridizes to a polyA region or vectorsequence, to identify sequences that are 5′ and 3′ of a known sequence.Additional techniques include capture PCR and walking PCR. Other methodsemploying amplification may also be employed to obtain a full lengthcDNA sequence.

[0069] In certain instances, it is possible to obtain a full length cDNAsequence by analysis of sequences provided in an expressed sequence tag(EST) database, such as that available from GenBank. Searches foroverlapping ESTs may generally be performed using well known programs(e.g., BLAST searches), and such ESTs may be used to generate acontiguous full length sequence. Full length DNA sequences may also beobtained by analysis of genomic fragments.

[0070] Polynucleotide variants may generally be prepared by any methodknown in the art, including chemical synthesis by, for example, solidphase phosphoramidite chemical synthesis. Modifications in apolynucleotide sequence may also be introduced using standardmutagenesis techniques, such as oligonucleotide-directed site-specificmutagenesis. Alternatively, RNA molecules may be generated by in vitroor in vivo transcription of DNA sequences encoding a breast tumorprotein, or portion thereof, provided that the DNA is incorporated intoa vector with a suitable RNA polymerase promoter (such as T7 or SP6).Certain portions may be used to prepare an encoded polypeptide, asdescribed herein. In addition a portion may be administered to a patientsuch that the encoded polypeptide is generated in vivo (e.g., bytransfecting antigen-presenting cells, such as dendritic cells, with acDNA construct encoding a breast tumor polypeptide, and administeringthe transfected cells to the patient).

[0071] A portion of a sequence complementary to a coding sequence (i.e.,an antisense polynucleotide) may also be used as a probe or to modulategene expression. cDNA constructs that can be transcribed into antisenseRNA may also be introduced into cells or tissues to facilitate theproduction of antisense RNA. An antisense polynucleotide may be used, asdescribed herein, to inhibit expression of a BCSG protein. Antisensetechnology can be used to control gene expression through triple-helixformation, which compromises the ability of the double helix to opensufficiently for the binding of polymerases, transcription factors orregulatory molecules. Alternatively, an antisense molecule may bedesigned to hybridize with a control region of a gene (e.g., promoter,enhancer or transcription initiation site), and block transcription ofthe gene; or to block translation by inhibiting binding of a transcriptto ribosomes.

[0072] A portion of a coding sequence, or of a complementary sequence,may also be designed as a probe or primer to detect gene expression.Probes may be labeled with a variety of reporter groups, such asradionuclides and enzymes, and are preferably at least 10 nucleotides inlength, more preferably at least 20 nucleotides in length and still morepreferably at least 30 nucleotides in length. Primers, as noted above,are preferably 22-30 nucleotides in length.

[0073] Any polynucleotide may be further modified to increase stabilityin vivo. Possible modifications include, but are not limited to, theaddition of flanking sequences at the 5′ and/or 3′ ends; the use ofphosphorothioate or 2-O-methyl rather than phosphodiesterase linkages inthe backbone; and/or the inclusion of nontraditional bases such asinosine, queosine and wybutosine, as well as acetyl- methyl-, thio- andother modified forms of adenine, cytidine, guanine, thymine and uridine.

[0074] Within certain embodiments, polynucleotides may be formulated soas to permit entry into a cell of a mammal, and expression therein. Suchformulations are particularly useful for therapeutic purposes, asdescribed below. Those of ordinary skill in the art will appreciate thatthere are many ways to achieve expression of a polynucleotide in atarget cell, and any suitable method may be employed. For example, apolynucleotide may be incorporated into a viral vector such as, but notlimited to, adenovirus, adeno-associated virus, retrovirus, or vacciniaor other pox virus (e.g., avian pox virus). The polynucleotides may alsobe administered as naked plasmid vectors. Techniques for incorporatingDNA into such vectors are well known to those of ordinary skill in theart.

[0075] Other formulations for therapeutic purposes include colloidaldispersion systems, such as macromolecule complexes, nanocapsules,microspheres, beads, and lipid-based systems including oil-in-wateremulsions, micelles, mixed micelles, and liposomes. A preferredcolloidal system for use as a delivery vehicle in vitro and in vivo is aliposome (i.e., an artificial membrane vesicle). The preparation and useof such systems is well known in the art.

[0076] BCSG-Related Polypeptides

[0077] Within the context of the present invention, BCSG-relatedpolypeptides comprise at least a biologically active portion or animmunogenic portion of a BCSG encoded polypeptide or a variant thereof.

[0078] Immunogenic portions may generally be identified using well knowntechniques. Such techniques include screening polypeptides for theability to react with antigen-specific antibodies, antisera and/orT-cell lines or clones. As used herein, antisera and antibodies are“antigen-specific” if they show immunospecific binding to an antigen(i.e., binding to the antigen with an affinity that is at least 10⁵M⁻¹).Such antisera and antibodies may be prepared as described herein, andusing well known techniques. An immunogenic portion of a native breastcancer protein is a portion that reacts with such antisera and/orT-cells at a level that is not substantially less than the reactivity ofthe full length polypeptide (e.g., in an ELISA and/or T-cell reactivityassay). Such immunogenic portions may react within such assays at alevel that is similar to or greater than the reactivity of the fulllength polypeptide. Such screens may generally be performed usingmethods well known to those of ordinary skill in the art, such as thosedescribed in Harlow and Lane, Antibodies: A Laboratory Manual, ColdSpring Harbor Laboratory, 1988. For example, a polypeptide may beimmobilized on a solid support and contacted with patient sera to allowbinding of antibodies within the sera to the immobilized polypeptide.Unbound sera may then be removed and bound antibodies detected using,for example, ¹²⁵I-labeled Protein A.

[0079] BCSG related polypeptides may comprise a signal (or leader)sequence at the N-terminal end of the protein, which co-translationallyor post-translationally directs transfer of the protein. The polypeptidemay also be conjugated to a linker or other sequence for ease ofsynthesis, purification or identification of the polypeptide (e.g.,poly-His), or to enhance binding of the polypeptide to a solid support.For example, a polypeptide may be conjugated to an immunoglobulin Fcregion.

[0080] BCSG related polypeptides may be prepared using any of a varietyof well known techniques. Recombinant polypeptides encoded bypolynucleotides as described above may be readily prepared from thepolynucleotides using any of a variety of expression vectors known tothose of ordinary skill in the art. Expression may be achieved in anyappropriate host cell that has been transformed or transfected with anexpression vector containing a DNA molecule that encodes a recombinantpolypeptide. Suitable host cells include prokaryotes, yeast, and highereukaryotic cells, such as mammalian cells and plant cells. Supernatantsfrom suitable host/vector systems which secrete recombinant protein orpolypeptide into culture media may be first concentrated using acommercially available filter. Following concentration, the concentratemay be applied to a suitable purification matrix such as an affinitymatrix or an ion exchange resin. Finally, one or more reverse phase HPLCsteps can be employed to further purify a recombinant polypeptide.

[0081] Portions and other variants having less than about 100 aminoacids, and generally less than about 50 amino acids, may also begenerated by synthetic means, using techniques well known to those ofordinary skill in the art. For example, such polypeptides may besynthesized using any of the commercially available solid-phasetechniques, such as the Merrifield solid-phase synthesis method, whereamino acids are sequentially added to a growing amino acid chain.Equipment for automated synthesis of polypeptides is commerciallyavailable from suppliers such as Perkin Elmer/Applied BioSystemsDivision (Foster City, Calif.), and may be operated according to themanufacturer's instructions.

[0082] Within certain specific embodiments, a polypeptide may be afusion protein that comprises multiple polypeptides as described herein,or that comprises at least one polypeptide as described herein and afusion partner. Certain preferred fusion partners are both immunologicaland expression enhancing fusion partners. Other fusion partners may beselected so as to increase the solubility of the protein or to enablethe protein to be targeted to desired intracellular compartments. Stillfurther fusion partners include affinity tags, which facilitatepurification of the protein.

[0083] Fusion proteins may generally be prepared using standardtechniques, including chemical conjugation. Preferably, a fusion proteinis expressed as a recombinant protein, allowing the production ofincreased levels, relative to a non-fused protein, in an expressionsystem. Briefly, DNA sequences encoding the polypeptide components maybe assembled separately, and ligated into an appropriate expressionvector. The 3′ end of the DNA sequence encoding one polypeptidecomponent is ligated, with or without a peptide linker, to the 5′ end ofa DNA sequence encoding the second polypeptide component so that thereading frames of the sequences are in phase. This permits translationinto a single fusion protein that retains the biological activity ofboth component polypeptides.

[0084] A peptide linker sequence may be employed to separate the firstand second polypeptide components by a distance sufficient to ensurethat each polypeptide folds into its secondary and tertiary structures.Such a peptide linker sequence is incorporated into the fusion proteinusing standard techniques well known in the art. Suitable peptide linkersequences may be chosen based on the following factors: (1) theirability to adopt a flexible extended conformation; (2) their inabilityto adopt a secondary structure that could interact with functionalepitopes on the first and second polypeptides; and (3) the lack ofhydrophobic or charged residues that might react with the polypeptidefunctional epitopes. Preferred peptide linker sequences contain Gly, Asnand Ser residues. Other near neutral amino acids, such as Thr and Alamay also be used in the linker sequence. Amino acid sequences which maybe usefully employed as linkers include those disclosed U.S. Pat. No.4,935,233 and U.S. Pat. No. 4,751,180. The linker sequence may generallybe from 1 to about 50 amino acids in length. Linker sequences are notrequired when the first and second polypeptides have non-essentialN-terminal amino acid regions that can be used to separate thefunctional domains and prevent steric interference.

[0085] The ligated DNA sequences are operably linked to suitabletranscriptional or translational regulatory elements. The regulatoryelements responsible for expression of DNA are located only 5′ to theDNA sequence encoding the first polypeptides. Similarly, stop codonsrequired to end translation and transcription termination signals areonly present 3′ to the DNA sequence encoding the second polypeptide.

[0086] Antibodies

[0087] The present invention further provides antibodies andantigen-binding fragments thereof, that specifically bind to a BCSM(BCSM-specific antibodies). As used herein, an antibody, orantigen-binding fragment thereof, is said to “specifically bind” to aBCSM if it binds to an antigen with an affinity that is at least 10⁵M⁻¹.As used herein, “binding” refers to a noncovalent association betweentwo separate molecules such that a complex is formed.

[0088] Antibodies may be prepared by any of a variety of techniquesknown to those of ordinary skill in the art. See, e.g., Harlow and Lane,Antibodies. A Laboratory Manual, Cold Spring Harbor Laboratory, 1988. Ingeneral, antibodies can be produced by cell culture techniques,including the generation of monoclonal antibodies as described herein,or via transfection of antibody genes into suitable bacterial ormammalian cell hosts, in order to allow for the production ofrecombinant antibodies. In one technique, an immunogen comprising thepolypeptide is initially injected into any of a wide variety of mammals(e.g., mice, rats, rabbits, sheep or goats). In this step, thepolypeptides of this invention may serve as the immunogen withoutmodification. Alternatively, particularly for relatively shortpolypeptides, a superior immune response may be elicited if thepolypeptide is joined to a carrier protein, such as bovine serum albuminor keyhole limpet hemocyanin. The immunogen is injected into the animalhost, preferably according to a predetermined schedule incorporating oneor more booster immunizations, and the animals are bled periodically.Polyclonal antibodies specific for the polypeptide may then be purifiedfrom such antisera by, for example, affinity chromatography using thepolypeptide coupled to a suitable solid support.

[0089] Monoclonal antibodies specific for an antigenic polypeptide ofinterest may be prepared, for example, using methods well known in theart. Briefly, these methods involve the preparation of immortal celllines capable of producing antibodies having the desired specificity(i.e., reactivity with the polypeptide of interest). Such cell lines maybe produced, for example, from spleen cells obtained from an animalimmunized as described above. The spleen cells are then immortalized by,for example, fusion with a myeloma cell fusion partner, preferably onethat is syngeneic with the immunized animal. A variety of fusiontechniques may be employed. For example, the spleen cells and myelomacells may be combined with a nonionic detergent for a few minutes andthen plated at low density on a selective medium that supports thegrowth of hybrid cells, but not myeloma cells. A preferred selectiontechnique uses HAT (hypoxanthine, aminopterin, thymidine) selection.After a sufficient time, usually about 1 to 2 weeks, breasties ofhybrids are observed. Single breasties are selected and their culturesupernatants tested for binding activity against the polypeptide.Hybridomas having high reactivity and specificity are preferred.

[0090] Monoclonal antibodies may be isolated from the supernatants ofgrowing hybridoma breasties. In addition, various techniques may beemployed to enhance the yield, such as injection of the hybridoma cellline into the peritoneal cavity of a suitable vertebrate host, such as amouse. Monoclonal antibodies may then be harvested from the ascitesfluid or the blood. Contaminants may be removed from the antibodies byconventional techniques, such as chromatography, gel filtration,precipitation, and extraction. The polypeptides of this invention may beused in the purification process in, for example, an affinitychromatography step.

[0091] Within certain embodiments, the use of antigen-binding fragmentsof antibodies may be preferred. Such fragments include Fab fragments,which may be prepared using standard techniques. Briefly,immunoglobulins may be purified from rabbit serum by affinitychromatography on Protein A bead columns and digested by papain to yieldFab and Fe fragments. The Fab and Fc fragments may be separated byaffinity chromatography on protein A bead columns.

[0092] Additionally, recombinant anti-BCSM antibodies, such as chimericand humanized monoclonal antibodies, comprising both human and non-humanportions, which can be made using standard recombinant DNA techniques,are within the scope of the invention. Such chimeric and humanizedmonoclonal antibodies can be produced by recombinant DNA techniquesknown in the art.

[0093] Humanized antibodies are particularly desirable for therapeutictreatment of human subjects. Humanized forms of non-human (e.g., murine)antibodies are chimeric molecules of immunoglobulins, immunoglobulinchains or fragments thereof (such as Fv, Fab, Fab′, F(ab′)₂ or otherantigen-binding subsequences of antibodies) which contain minimalsequence derived from non-human immunoglobulin. Humanized antibodiesinclude human immunoglobulins (recipient antibody) in which residuesforming a complementary determining region (CDR) of the recipient arereplaced by residues from a CDR of a non-human species (donor antibody)such as mouse, rat or rabbit having the desired specificity, affinityand capacity. In some instances, Fv framework residues of the humanimmunoglobulin are replaced by corresponding non-human residues.Humanized antibodies may also comprise residues which are found neitherin the recipient antibody nor in the imported CDR or frameworksequences. In general, the humanized antibody will comprisesubstantially all of at least one, and typically two, variable domains,in which all or substantially all of the CDR regions correspond to thoseof a non-human immunoglobulin and all or substantially all of theconstant regions being those of a human immunoglobulin consensussequence. The humanized antibody will preferably also comprise at leasta portion of an immunoglobulin constant region (Fe), typically that of ahuman immunoglobulin.

[0094] A therapeutic agent may be coupled (e.g., covalently bonded) to asuitable antibody either directly or indirectly (e.g., via a linkergroup). A direct reaction between an agent and an antibody is possiblewhen each possesses a substituent capable of reacting with the other.For example, a nucleophilic group, such as an amino or sulfhydryl group,on one may be capable of reacting with a carbonyl-containing group, suchas an anhydride or an acid halide, or with an alkyl group containing agood leaving group (e.g., a halide) on the other.

[0095] Alternatively, it may be desirable to couple a therapeutic agentand an antibody via a linker group. A linker group can function as aspacer to distance an antibody from an agent in order to avoidinterference with binding capabilities. A linker group can also serve toincrease the chemical reactivity of a substituent on an agent or anantibody, and thus increase the coupling efficiency. An increase inchemical reactivity may also facilitate the use of agents, or functionalgroups on agents, which otherwise would not be possible.

[0096] It may be desirable to couple more than one agent to an antibody.In one embodiment, multiple molecules of an agent are coupled to oneantibody molecule. In another embodiment, more than one type of agentmay be coupled to one antibody. Regardless of the particular embodiment,immunoconjugates with more than one agent may be prepared in a varietyof ways. For example, more than one agent may be coupled directly to anantibody molecule, or linkers that provide multiple sites for attachmentcan be used.

[0097] Vectors

[0098] Another aspect of the invention pertains to vectors containing apolynucleotide encoding a BCSG protein, or a portion thereof. One typeof vector is a “plasmid,” which includes a circular double stranded DNAloop into which additional DNA segments can be ligated. Vectors includeexpression vectors and gene delivery vectors.

[0099] The expression vectors of the invention comprise a polynucleotideencoding a BCSG protein or a portion thereof in a form suitable forexpression of the polynucleotide in a host cell, which means that theexpression vectors include one or more regulatory sequences, selected onthe basis of the host cells to be used for expression, which isoperatively linked to the polynucleotide sequence to be expressed. Itwill be appreciated by those skilled in the art that the design of theexpression vector can depend on such factors as the choice of the hostcell to be transformed, the level of expression of protein desired, andthe like. The expression vectors of the invention can be introduced intohost cells to thereby produce proteins or peptides, including fusionproteins or peptides, encoded by polynucleotides as described herein(e.g., BCSG polypeptides, variants of BCSG polypeptides, fusionproteins, and the like).

[0100] The expression vectors of the invention can be designed forexpression of BCSG polypeptides in prokaryotic or eukaryotic cells. Forexample, BCSG polypeptides can be expressed in bacterial cells such asE. coli, insect cells (using baculovirus expression vectors) yeast cellsor mammalian cells. In certain embodiments, such protein may be used,for example, as a therapeutic protein of the invention. Alternatively,the expression vector can be transcribed and translated in vitro, forexample using T7 promoter regulatory sequences and T7 polymerase.

[0101] In another embodiment, the expression vector is a yeastexpression vector. Examples of vectors for expression in yeast S.cerevisiae include pYepSec1, pMFa, pJRY88, pYES2 (InvitrogenCorporation, San Diego, Calif.), and picZ (Invitrogen Corp, San Diego,Calif.).

[0102] Alternatively, BCSG polypeptides of the invention can beexpressed in insect cells using baculovirus expression vectors.Baculovirus vectors available for expression of proteins in culturedinsect cells (e.g., Sf9 cells) include the pAc series and the pVLseries.

[0103] In yet another embodiment, a BCSG is expressed in mammalian cellsusing a mammalian expression vector. When used in mammalian cells, theexpression vector's control functions are often provided by viralregulatory elements. For example, commonly used promoters are derivedfrom polyoma, adenovirus 2, cytomegalovirus and Simian Virus 40.

[0104] In another embodiment, the mammalian expression vector is capableof directing expression of the polynucleotide preferentially in aparticular cell type (e.g., tissue-specific regulatory elements are usedto express the polynucleotide). Tissue-specific regulatory elements areknown in the art and may include epithelial cell-specific promoters.Other non-limiting examples of suitable tissue-specific promotersinclude the liver-specific albumin promoter, lymphoid-specificpromoters, promoters of T cell receptors and immunoglobulins,neuron-specific promoters (e.g., the neurofilament promoter),pancreas-specific promoters, and mammary gland-specific promoters (e.g.,milk whey promoter). Developmentally-regulated promoters are alsoencompassed, for example the marine box promoters and the α-fetoproteinpromoter. In certain preferred embodiments of the invention, thetissue-specific promoter is an epithelial cell-specific promoter.

[0105] The invention provides a recombinant expression vector comprisinga polynucleotide encoding a BCSG cloned into the expression vector in anantisense orientation. That is, the DNA molecule is operatively linkedto a regulatory sequence in a manner which allows for expression (bytranscription of the DNA molecule) of an RNA molecule which is antisenseto mRNA corresponding to a BCSG of the invention. Regulatory sequencesoperatively linked to a polynucleotide cloned in the antisenseorientation can be chosen which direct the continuous expression of theantisense RNA molecule in a variety of cell types, for instance viralpromoters and/or enhancers, or regulatory sequences can be chosen whichdirect constitutive, tissue specific or cell type specific expression ofantisense RNA. The antisense expression vector can be in the form of arecombinant plasmid, phagemid or attenuated virus in which antisensepolynucleotides are produced under the control of a high efficiencyregulatory region, the activity of which can be determined by the celltype into which the vector is introduced.

[0106] The invention further provides gene delivery vectors for deliveryof polynucleotides to cells, tissue, or to a the mammal for expression.For example, a polynucleotide sequence of the invention can beadministered either locally or systemically in a gene delivery vector.These constructs can utilize viral or non-viral vector approaches in invivo or ex vivo modality. Expression of such coding sequence can beinduced using endogenous mammalian or heterologous promoters. Expressionof the coding sequence in vivo can be either constituted or regulated.The invention includes gene delivery vehicles capable of expressing thecontemplated polynucleotides. The gene delivery vehicle is preferably aviral vector and, more preferably, a retroviral, lentiviral, adenoviral,adeno-associated viral (AAV), herpes viral, or alphavirus vectors. Theviral vector can also be an astrovirus, coronavirus, orthomyxovirus,papovavirus, paramyxovirus, parvovirus, picornavirus, poxvirus,togavirus viral vector.

[0107] Delivery of the gene therapy constructs of this invention intocells is not limited to the above mentioned viral vectors. Otherdelivery methods and media may be employed such as, for example,liposomes, polycationic condensed DNA linked or unlinked to inactivatedadenovirus, ligand linked DNA, naked DNA and eucaryotic cell deliveryvehicles cells.

[0108] Another aspect of the invention pertains to the expression ofBCSGs using a regulatable expression system. Systems to regulateexpression of therapeutic genes have been developed and incorporatedinto the current viral and nonviral gene delivery vectors. Examples ofregulatable systems include: the tet-on/off system, the ecdysone system,the progesterone-system, and the rapamycin system.

[0109] Methods for Detecting Breast Cancer

[0110] In general, breast cancer may be detected in a patient based onthe presence of one or more BCSG products (polynucleotides orpolypeptide) in a biological sample (for example, blood, sera, sputumurine and/or tumor biopsies) obtained from the patient. In other words,such BCSG products may be used as markers to indicate the presence orabsence of breast cancer. In addition, such products may be useful forthe detection of other cancers. The antibodies provided herein generallypermit detection of the level of antigen that binds to the agent in thebiological sample. Polynucleotide primers and probes may be used todetect the levels of transcribed polynucleotides from BCSGs, which isalso indicative of the presence or absence of a cancer.

[0111] There are a variety of assay formats known to those of ordinaryskill in the art for using an antibody to detect polypeptide markers ina sample. See, e.g., Harlow and Lane, Antibodies: A Laboratory Manual,Cold Spring Harbor Laboratory, 1988. In general, the presence or absenceof a cancer in a patient may be determined by (a) contacting abiological sample obtained from a patient with an antibody; (b)detecting in the sample a level of polypeptide that binds to theantibody; and (c) comparing the level of polypeptide with apredetermined control value.

[0112] In a preferred embodiment, the assay involves the use of antibodyimmobilized on a solid support to bind to and remove the polypeptidefrom the remainder of the sample. The bound polypeptide may then bedetected using a detection reagent that contains a reporter group andspecifically binds to the antibody/polypeptide complex. Such detectionreagents may comprise, for example, an antibody that specifically bindsto the polypeptide or an antibody or other agent that specifically bindsto the antibody, such as an anti-immunoglobulin, protein G, protein A ora lectin. Alternatively, a competitive assay may be utilized, in which apolypeptide is labeled with a reporter group and allowed to bind to theimmobilized antibody after incubation of the antibody with the sample.The extent to which components of the sample inhibit the binding of thelabeled polypeptide to the antibody is indicative of the reactivity ofthe sample with the immobilized antibody. Suitable polypeptides for usewithin such assays include full length breast tumor proteins andportions thereof to which the antibody binds, as described above.

[0113] The solid support may be any material known to those of ordinaryskill in the art to which the tumor protein may be attached. Forexample, the solid support may be a test well in a microtiter plate or anitrocellulose or other suitable membrane. Alternatively, the supportmay be a bead or disc, such as glass, fiberglass, latex or a plasticmaterial such as polystyrene or polyvinylchloride. The support may alsobe a magnetic particle or a fiber optic sensor, such as those disclosed,for example, in U.S. Pat. No. 5,359,681. The antibody may be immobilizedon the solid support using a variety of techniques known to those ofskill in the art. In the context of the present invention, the term“immobilization” refers to both noncovalent association, such asadsorption, and covalent attachment (which may be a direct linkagebetween the antibody and functional groups on the support or may be alinkage by way of a cross-linking agent). Immobilization by adsorptionto a well in a microtiter plate or to a membrane is preferred. In suchcases, adsorption may be achieved by contacting the antibody, in asuitable buffer, with the solid support for a suitable amount of time.The contact time varies with temperature, but is typically between about1 hour and about 1 day. In general, contacting a well of a plasticmicrotiter plate (such as polystyrene or polyvinylchloride) with anamount of antibody ranging from about 10 ng to about 10 μg, andpreferably about 100 ng to about 1 μg, is sufficient to immobilize anadequate amount of the antibody.

[0114] In certain embodiments, the assay is a two-antibody sandwichassay. This assay may be performed by first contacting an antibody thathas been immobilized on a solid support, commonly the well of amicrotiter plate, with the sample, such that polypeptides within thesample are allowed to bind to the immobilized antibody. Unbound sampleis then removed from the immobilized polypeptide-antibody complexes anda detection reagent (preferably a second antibody capable of binding toa different site on the polypeptide) containing a reporter group isadded. The amount of detection reagent that remains bound to the solidsupport is then determined using a method appropriate for the specificreporter group.

[0115] To determine the presence or absence of breast cancer, the signaldetected from the reporter group that remains bound to the solid supportis generally compared to a signal that corresponds to a predeterminedcontrol value. In one preferred embodiment, the control value for thedetection of breast cancer is the average mean signal obtained when theimmobilized antibody is incubated with samples from patients without thecancer. A sample generating a signal that is significantly higher (e.g.,≧200%) or lower (e.g., ≦50%) than the control value determined by thismethod may be considered indicative of cancer.

[0116] In a related embodiment, the assay is performed in a flow-throughor strip test format, wherein the antibody is immobilized on a membrane,such as nitrocellulose. In the flow-through test, polypeptides withinthe sample bind to the immobilized binding agent as the sample passesthrough the membrane. A second, labeled binding agent then binds to thebinding agent-polypeptide complex as a solution containing the secondbinding agent flows through the membrane. The detection of bound secondbinding agent may then be performed as described above. In the striptest format, one end of the membrane to which binding agent is bound isimmersed in a solution containing the sample. The sample migrates alongthe membrane through a region containing second binding agent and to thearea of immobilized binding agent. Concentration of second binding agentat the area of immobilized antibody indicates the presence of a cancer.Typically, the concentration of second binding agent at that sitegenerates a pattern, such as a line, that can be read visually. Theabsence of such a pattern indicates a negative result. In general, theamount of binding agent immobilized on the membrane is selected togenerate a visually discernible pattern when the biological samplecontains a level of polypeptide that would be sufficient to generate apositive signal in the two-antibody sandwich assay, in the formatdiscussed above. Preferred binding agents for use in such assays areantibodies and antigen-binding fragments thereof. Preferably, the amountof antibody immobilized on the membrane ranges from about 25 ng to about1 μg, and more preferably from about 50 ng to about 500 ng. Such testscan typically be performed with a very small amount of biologicalsample.

[0117] Numerous other assay protocols exist that are suitable for usewith the BCSG products or antibodies of the present invention. The abovedescriptions are intended to be exemplary only. For example, it will beapparent to those of ordinary skill in the art that the above protocolsmay be readily modified to use BCSG polypeptides to detect antibodiesthat bind to such polypeptides in a biological sample. The detection ofsuch BCSG-specific antibodies may correlate with the presence of breastcancer.

[0118] As noted above, breast cancer may also, or alternatively, bedetected based on the level of mRNA transcribed from a BCSG in abiological sample. For example, at least two oligonucleotide primers maybe employed in a polymerase chain reaction (PCR) based assay to amplifya portion of a breast tumor cDNA derived from a biological sample,wherein at least one of the oligonucleotide primers is specific for(i.e., hybridizes to) a polynucleotide encoding the breast tumorprotein. The amplified cDNA is then separated and detected usingtechniques well known in the art, such as gel electrophoresis.Similarly, oligonucleotide probes that specifically hybridize to apolynucleotide encoding a breast tumor protein may be used in ahybridization assay to detect the presence of polynucleotide encodingthe tumor protein in a biological sample.

[0119] To permit hybridization under assay conditions, oligonucleotideprimers and probes should comprise an oligonucleotide sequence that hasat least about 70%, preferably at least about 80% and more preferably atleast about 90%, identity to a portion of a polynucleotide encoding abreast tumor protein that is at least 10 nucleotides, and preferably atleast 20 nucleotides, in length. Preferably, oligonucleotide primersand/or probes hybridize to a polynucleotide encoding a polypeptidedescribed herein under moderately stringent conditions, as definedabove. Oligonucleotide primers and/or probes which may be usefullyemployed in the diagnostic methods described herein preferably are atleast 10-40 nucleotides in length. In a preferred embodiment, theoligonucleotide primers comprise at least 10 contiguous nucleotides,more preferably at least 15 contiguous nucleotides, of a DNA moleculehaving a sequence recited in SEQ ID NOS:1-19. Techniques for both PCRbased assays and hybridization assays are well known in the art.

[0120] One preferred assay employs RT-PCR, in which PCR is applied inconjunction with reverse transcription. Typically, RNA is extracted froma biological sample, such as biopsy tissue, and is reverse transcribedto produce cDNA molecules. PCR amplification using at least one specificprimer generates a cDNA molecule, which may be separated and visualizedusing, for example, gel electrophoresis. Amplification may be performedon biological samples taken from a test patient and from an individualwho is not afflicted with a cancer. The amplification reaction may beperformed on several dilutions of cDNA spanning two orders of magnitude.A two-fold or greater increase/decrease in expression in severaldilutions of the test patient sample as compared to the same dilutionsof the non-cancerous sample may be considered indicative of cancer.

[0121] As noted above, to improve sensitivity, multiple BCSG markers maybe assayed within a given sample. It will be apparent that antibodiesspecific for different proteins provided herein may be combined within asingle assay. Further, multiple primers or probes may be usedconcurrently. The selection of BCSG markers may be based on routineexperiments to determine combinations that results in optimalsensitivity. In addition, or alternatively, assays for BCSG productsprovided herein may be combined with assays for other known tumorantigens.

[0122] Diagnostic Kits

[0123] The present invention further provides kits for use within any ofthe above diagnostic methods. Such kits typically comprise two or morecomponents necessary for performing a diagnostic assay. Components maybe compounds, reagents, containers and/or equipment. For example, onecontainer within a kit may contain a monoclonal antibody or fragmentthereof that specifically binds to a polypeptide. Such antibodies orfragments may be provided attached to a support material, as describedabove. One or more additional containers may enclose elements, such asreagents or buffers, to be used in the assay. Such kits may also, oralternatively, contain a detection reagent as described above thatcontains a reporter group suitable for direct or indirect detection ofantibody binding.

[0124] Alternatively, a kit may contain at least one oligonucleotideprobe or primer, as described above, that hybridizes to a polynucleotidetranscribed from a BCSG. Such an oligonucleotide may be used, forexample, within a PCR or hybridization assay. Additional components thatmay be present within such kits include a second oligonucleotide and/ora diagnostic reagent or container to facilitate the detection of apolynucleotide transcribed from a BCSG.

[0125] Arrays and Biochips

[0126] The invention also includes an array comprising a panel of BCSMsof the present invention. The array can be used to assay expression ofone or more genes in the array.

[0127] It will be appreciated by one skilled in the art that the panelsof BCSMs of the invention may conveniently be provided on solidsupports, as a biochip. For example, polynucleotides may be coupled toan array (e.g., a biochip using GeneChip® for hybridization analysis),to a resin (e.g., a resin which can be packed into a column for columnchromatography), or a matrix (e.g., a nitrocellulose matrix for northernblot analysis). The immobilization of molecules complementary to theBCSG(s), either covalently or noncovalently, permits a discrete analysisof the presence or activity of each BCSG in a sample. In an array, forexample, polynucleotides complementary to each member of a panel ofBCSGs may individually be attached to different, known locations on thearray. The array may be hybridized with, for example, polynucleotidesextracted from a blood or colon sample from a subject. The hybridizationof polynucleotides from the sample with the array at any location on thearray can be detected, and thus the presence or quantity of the BCSG andBCSG transcripts in the sample can be ascertained. In a preferredembodiment, an array based on a biochip is employed. Similarly, Westernanalyses may be performed on immobilized antibodies specific for BCSMshybridized to a protein sample from a subject.

[0128] It will also be apparent to one skilled in the art that theentire BCSM (protein or polynucleotide) molecule need not be conjugatedto the biochip support; a portion of the BCSM or sufficient length fordetection purposes (i.e., for hybridization), for example a portion ofthe BCSM which is 7, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70,75, 100 or more nucleotides or amino acids in length may be sufficientfor detection purposes.

[0129] Identifying Modulators of BCSM

[0130] The invention also provides methods for identifying modulators,i.e., candidate agents which (a) bind to a BCSM, or (b) have amodulatory (e.g., stimulatory or inhibitory) effect on the activity of aBCSM or, more specifically, (c) have a modulatory effect on theinteractions of the BCSM with one or more of its natural substrates(e.g., peptide, protein, hormone, co-factor, or polynucleotide), or (d)have a modulatory effect on the expression of the BCSMs. Such assaystypically comprise a reaction between the BCSM and one or more assaycomponents. The other components may be either the candidate agentsitself, or a combination of candidate agents and a binding partner ofthe BCSM.

[0131] The candidate agents of the present invention are generallyeither small molecules or bioactive agents. In one embodiment the testcompound is a small molecule. In another embodiment, the test compoundis a bioactive agent. Bioactive agents include but are not limited tonaturally-occurring or synthetic compounds or biomolecules. One skilledin the art will appreciate that the nature of the candidate agents mayvary depending on the nature of the protein encoded by the BCSG of theinvention. For example, if the BCSG encodes an orphan receptor having anunknown ligand, the test compound may be any of a number of bioactiveagents which may act as cognate ligand, including but not limited to,cytokines, lipid-derived mediators, small biogenic amines, hormones,neuropeptides, or proteases. In another embodiment, the candidate agentscan be an antisense polynucleotide molecule which is complementary to aBCSG polynucleotides.

[0132] As used herein, the term “binding partner” refers to a bioactiveagent which serves as either a substrate for a BCSM, or alternatively,as a ligand having binding affinity to the BCSM.

[0133] Modulators of BCSG expression, activity or binding ability areuseful as thereapeutic compositions of the invention. Such modulators(e.g., antagonists or agonists) may be formulated as pharmaceuticalcompositions, as described herein below. Such modulators may also beused in the methods of the invention, for example, to diagnose, treat,or prognose breast cancer.

[0134] Vaccines

[0135] Within certain aspects, BCSG products (polypeptides andpolynucleotides) described herein may be used as vaccines for breastcancer. Vaccines may comprise one or more such products and animmunostimulant. An immunostimulant may be any substance that enhancesor potentiates an immune response (antibody and/or cell-mediated) to anexogenous antigen. Examples of immunostimulants include adjuvants,biodegradable microspheres (e.g., polylactic galactide) and liposomes.Vaccines within the scope of the present invention may also containother compounds, which may be biologically active or inactive. Forexample, one or more immunogenic portions of other tumor antigens may bepresent, either incorporated into a fusion polypeptide or as a separatecompound, within the composition or vaccine.

[0136] A vaccine may contain DNA encoding one or more of thepolypeptides as described above, such that the polypeptide is generatedin situ. As noted above, the DNA may be present within any of a varietyof delivery systems known to those of ordinary skill in the art,including nucleic acid expression systems, bacteria and viral expressionsystems. Numerous gene delivery techniques are well known in the art.Appropriate nucleic acid expression systems contain the necessary DNAsequences for expression in the patient (such as a suitable promoter andterminating signal). Bacterial delivery systems involve theadministration of a bacterium (such as Bacillus-Calmette-Guerrin) thatexpresses an immunogenic portion of the polypeptide on its cell surfaceor secretes such an epitope. In a preferred embodiment, the DNA may beintroduced using a viral expression system (e.g., vaccinia or other poxvirus, retrovirus, or adenovirus), which may involve the use of anon-pathogenic (defective), replication competent virus. Techniques forincorporating DNA into such expression systems are well known to thoseof ordinary skill in the art. The DNA may also be naked DNA. The uptakeof naked DNA may be increased by coating the DNA onto biodegradablebeads, which are efficiently transported into the cells. It will beapparent that a vaccine may comprise both a polynucleotide and apolypeptide component. Such vaccines may provide for an enhanced immuneresponse.

[0137] It will be apparent that a vaccine may contain pharmaceuticallyacceptable salts of the polynucleotides and polypeptides providedherein. Such salts may be prepared from pharmaceutically acceptablenon-toxic bases, including organic bases (e.g., salts of primary,secondary and tertiary amines and basic amino acids) and inorganic bases(e.g., sodium, potassium, lithium, ammonium, calcium and magnesiumsalts).

[0138] Any of a variety of immunostimulants may be employed in thevaccines of this invention. For example, an adjuvant may be included.Most adjuvants contain a substance designed to protect the antigen fromrapid catabolism, such as aluminum hydroxide or mineral oil, and astimulator of immune responses, such as lipid A, Bortadellci pertussisor Mycobacterium tuberculosis derived proteins. Suitable adjuvants arecommercially available as, for example, Freund's Incomplete Adjuvant andComplete Adjuvant (Difco Laboratories, Detroit, Mich.); Merck Adjuvant65 (Merck and Company, Inc., Rahway, N.J.); AS-2 (SmithKline Beecham,Philadelphia, Pa.); aluminum salts such as aluminum hydroxide gel (alum)or aluminum phosphate; salts of calcium, iron or zinc; an insolublesuspension of acylated tyrosine; acylated sugars; cationically oranionically derivatized polysaccharides; polyphosphazenes; biodegradablemicro spheres; monophosphoryl lipid A and quil A. Cytokines, such asGM-CSF or interleukin-2, -7, or -12, may also be used as adjuvants.

[0139] Any vaccine provided herein may be prepared using well knownmethods that result in a combination of antigen, immune responseenhancer and a suitable carrier or excipient. The compositions describedherein may be administered as part of a sustained release formulation(i.e., a formulation such as a capsule, sponge or gel (composed ofpolysaccharides, for example) that effects a slow release of compoundfollowing administration). Such formulations may generally be preparedusing well known technology and administered by, for example, oral,rectal or subcutaneous implantation, or by implantation at the desiredtarget site. Sustained-release formulations may contain a polypeptide,polynucleotide or antibody dispersed in a carrier matrix and/orcontained within a reservoir surrounded by a rate controlling membrane.

[0140] Carriers for use within such formulations are biocompatible, andmay also be biodegradable; preferably the formulation provides arelatively constant level of active component release. Such carriersinclude microparticles of poly(lactide-co-glycolide), as well aspolyacrylate, latex, starch, cellulose and dextran. Otherdelayed-release carriers include supramolecular biovectors, whichcomprise a non-liquid hydrophilic core (e.g., a cross-linkedpolysaccharide or oligosaccharide) and, optionally, an external layercomprising an amphiphilic compound, such as a phospholipid. The amountof active compound contained within a sustained release formulationdepends upon the site of implantation, the rate and expected duration ofrelease and the nature of the condition to be treated or prevented.

[0141] Pharmaceutical Compositions

[0142] The invention is further directed to pharmaceutical compositionscomprising a pharmaceutically acceptable carrier and at least one of thefollowing: a BCSM, a variant of a BCSM, a BCSM modulator, aBCSM-specific antibody, a vaccine generated using a BCSM or its variant,and a vector capable of expressing a BCSM or a variant of a BCSM.

[0143] As used herein the language “pharmaceutically acceptable carrier”is intended to include any and all solvents, solubilizers, fillers,stabilizers, binders, absorbents, bases, buffering agents, lubricants,controlled release vehicles, diluents, emulsifying agents, humectants,lubricants, dispersion media, coatings, antibacterial or antifungalagents, isotonic and absorption delaying agents, and the like,compatible with pharmaceutical administration. The use of such media andagents for pharmaceutically active substances is well-known in the art.Except insofar as any conventional media or agent is incompatible withthe active compound, use thereof in the compositions is contemplated.Supplementary agents can also be incorporated into the compositions.

[0144] The invention includes methods for preparing pharmaceuticalcompositions for modulating the expression or activity of a BCSM of theinvention. Such methods comprise formulating a pharmaceuticallyacceptable carrier with an agent which modulates expression or activityof a BCSM . Such compositions can further include additional activeagents. Thus, the invention further includes methods for preparing apharmaceutical composition by formulating a pharmaceutically acceptablecarrier with an agent which modulates expression or activity of a BCSMand one or more additional bioactive agents.

[0145] A pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. Examples of routesof administration include parenteral, e.g., intravenous, intradermal,subcutaneous, oral (e.g., inhalation), transdermal (topical),transmucosal, and rectal administration. Solutions or suspensions usedfor parenteral, intradermal, or subcutaneous application can include thefollowing components: a sterile diluent such as water for injection,saline solution, fixed oils, polyethylene glycols, glycerine; propyleneglycol or other synthetic solvents; antibacterial agents such as benzylalcohol or methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfate; chelating agents such as ethylenediaminetetraacetic acid;buffers such as acetates, citrates or phosphates and agents for theadjustment of tonicity such as sodium chloride or dextrose. pH can beadjusted with acids or bases, such as hydrochloric acid or sodiumhydroxide. The parenteral preparation can be enclosed in ampoules,disposable syringes or multiple dose vials made of glass or plastic.

[0146] Pharmaceutical compositions suitable for injectable use includesterile aqueous solutions (where water soluble) or dispersions andsterile powders for the extemporaneous preparation of sterile injectablesolutions or dispersion. For intravenous administration, suitablecarriers include physiological saline, bacteriostatic water, CremophorEL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In allcases, the injectable composition should be sterile and should be fluidto the extent that easy syringability exists. It must be stable underthe conditions of manufacture and storage and must be preserved againstthe contaminating action of microorganisms such as bacteria and fungi.The carrier can be a solvent or dispersion medium containing, forexample, water, ethanol, polyol (for example, glycerol, propyleneglycol, and liquid polyetheylene glycol, and the like), and suitablemixtures thereof. The proper fluidity can be maintained, for example, bythe use of a coating such as lecithin, by the maintenance of therequited particle size in the case of dispersion and by the use ofsurfactants. Prevention of the action of microorganisms can be achievedby various antibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars, polyalcohols such as manitol, sorbitol, sodium chloride in thecomposition. Prolonged absorption of the injectable compositions can bebrought about by including in the composition an agent which delaysabsorption, for example, aluminum monostearate and gelatin.

[0147] Sterile injectable solutions can be prepared by incorporating theactive compound (e.g., a fragment of a BCSM or an anti-BCSM antibody) inthe required amount in an appropriate solvent with one or a combinationof ingredients enumerated above, as required, followed by filteredsterilization. Generally, dispersions are prepared by incorporating theactive compound into a sterile vehicle which contains a basic dispersionmedium and the required other ingredients from those enumerated above.In the case of sterile powders for the preparation of sterile injectablesolutions, the preferred methods of preparation are vacuum drying andfreeze-drying which yields a powder of the active, ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

[0148] Oral compositions generally include an inert diluent or an ediblecarrier. They can be enclosed in gelatin capsules or compressed intotablets. For the purpose of oral therapeutic administration, the activecompound can be incorporated with excipients and used in the form oftablets, troches, or capsules. Oral compositions can also be preparedusing a fluid carrier for use as a mouthwash, wherein the compound inthe fluid carrier is applied orally and swished and expectorated orswallowed. Pharmaceutically compatible binding agents, and/or adjuvantmaterials can be included as part of the composition. The tablets,pills, capsules, troches and the like can contain any of the followingingredients, or compounds of a similar nature: a binder such asmicrocrystalline cellulose, gum tragacanth or gelatin; an excipient suchas starch or lactose, a disintegrating agent such as alginic acid,Primogel, or corn starch; a lubricant such as magnesium stearate orStertes; a glidant such as colloidal silicon dioxide; a sweetening agentsuch as sucrose or saccharin; or a flavoring agent such as peppermint,methyl salicylate, or orange flavoring.

[0149] For administration by inhalation, the compounds are delivered inthe form of an aerosol spray from pressured container or dispenser whichcontains a suitable propellant, e.g., a gas such as carbon dioxide, or anebulizer.

[0150] Systemic administration can also be by transmucosal ortransdermal means. For transmucosal or transdermal administration,penetrants appropriate to the barrier to be permeated are used in theformulation. Such penetrants are generally known in the art, andinclude, for example, for transmucosal administration, detergents, bilesalts, and fusidic acid derivatives. Transmucosal administration can beaccomplished through the use of nasal sprays or suppositories. Fortransdermal administration, the bioactive compounds are formulated intoointments, salves, gels, or creams as generally known in the art.

[0151] In one embodiment, the therapeutic moieties, which may contain abioactive compound, are prepared with carriers that will protect thecompound against rapid elimination from the body, such as a controlledrelease formulation, including implants and microencapsulated deliverysystems. Biodegradable, biocompatible polymers can be used, such asethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters, and polylactic acid. Methods for preparation of suchformulations will be apparent to those skilled in the art. The materialscan also be obtained commercially from e.g. Alza Corporation and NovaPharmaceuticals, Inc. Liposomal suspensions (including liposomestargeted to infected cells with monoclonal antibodies to viral antigens)can also be used as pharmaceutically acceptable carriers.

[0152] It is especially advantageous to formulate oral or parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein includesphysically discrete units suited as unitary dosages for the subject tobe treated; each unit containing a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on the unique characteristics of the active compound and theparticular therapeutic effect to be achieved, and the limitationsinherent in the art of compounding such an active compound for thetreatment of individuals.

[0153] The BCSGs of the invention can be inserted into gene deliveryvectors and used as gene therapy vectors. Gene therapy vectors can bedelivered to a subject by intravascular, intrameucular, subcutaneous,intraperitoneal injection, by direct injection into the target tissue,by inhalation, or by perfusion. The pharmaceutical preparation of thegene therapy vector can include the gene therapy vector in an acceptablediluent, or can comprise a slow release matrix in which the genedelivery vehicle is imbedded. Alternatively, where the complete genedelivery vector can be produced intact from recombinant cells, e.g.,retroviral vectors, the pharmaceutical preparation can include one ormore cells which produce the gene delivery system.

[0154] The pharmaceutical compositions can be included in a container,pack, or dispenser together with instructions for administration.

[0155] Methods for Treating Breast Cancer

[0156] In further aspects of the present invention, the pharmaceuticalcompositions described herein may be used for treatment of breastcancer. Within such methods, pharmaceutical compositions are typicallyadministered to a patient. A patient may or may not be afflicted withcancer. Accordingly, the above pharmaceutical compositions may be usedto prevent the development of breast cancer or to treat a patientafflicted with breast cancer. Breast cancer may be diagnosed usingcriteria generally accepted in the art, including the detection methoddescribed herein. Pharmaceutical compositions may be administered eitherprior to or following surgical removal of primary tumors and/ortreatment such as administration of radiotherapy or conventionalchemotherapeutic drugs.

[0157] Routes and frequency of administration of the pharmaceuticalcompositions described herein, as well as dosage, will vary fromindividual to individual, and may be readily established using standardtechniques. In general, an appropriate dosage and treatment regimenprovides the pharmaceutical composition(s) in an amount sufficient toprovide therapeutic and/or prophylactic benefit. Such a response can bemonitored by establishing an improved clinical outcome (e.g., morefrequent remissions, complete or partial, or longer disease-freesurvival) in treated patients as compared to non-treated patients.

EXAMPLES

[0158] The following Examples are offered by way of illustration and notby way of limitation.

Example 1 Identification of Genes Differentially Expressed Between theMetastatic Breast Cancer Cell Line MDA-MB-231 and the Non-TumorigenicDerivative MDA/H6 Using High Density Gene Filters

[0159] Total RNA was extracted from MDA-MB-231 and MDA/H6 cells withTrizol Reagent (15596-026, Life Technologies, Rockville, Md.) followingthe manufacturer's instructions. Briefly, cells were lysed by adding17.5 ml Trizol solution per 175 cm² flask. After transferring the lysateinto a tube, 0.2 ml chloroform was added per 1 ml Trizol reagent used.The samples were centrifuged at 12,000 g for 15 min at 4° C. The aqueousphase was transferred to a fresh tube, and 0.9 ml isopropyl alcohol wasadded each ml of aqueous phase collected. The samples were incubated atroom temperature for 10 min and spun at 12,000 g for 10 min at 4° C. Thesupernatant was removed and the RNA pellet was washed once with 75%ethanol alcohol. The pellet was air-dried and then dissolved inRNase-free water (D-5758, Sigma, St. Louis, Mo.). RNA was purified usingRneasy Midi Kit 50 (75144, Qiagen, Valencia, Calif.) followingmanufacturer's instructions. Briefly, 500 μg total RNA was purified byuse of 1 mg purification column. The RNA was equalized to 1 mlRnase-free water and then 3.8 ml Buffer RLT was added. Next, 2.8 ml 100%ethanol alcohol was added and the sample was placed on the Rneasy midispin column. The column was centrifuged for 5 min at 5,000 g, and theflow-through was discarded. Two and a 0.5 ml Buffer RPE was added to thecolumn that was centrifuged at 5,000 g for 5 min., and repeated once.The column was transferred to a new collection tube and 250 μlRNase-free water was added to the column and spun at 5,000 g for 5 min.This elution step was repeated once. Both of the elution weretransferred into a Microcon 100 column and spun at 500 g for 12 min. Thecolumn was inversely placed into a tube and spun at 3,000 g for 3 min.to collect the concentrated RNA.

[0160] High density gene filters (gf200, gf201, gf202, gf203 and gf211)consisting of 25,985 arrayed elements (19,592 unique human genes and6,393 controls) were purchased from Research Genetics (Huntsville,Ala.). A new gene filter was first washed in boiling 0.5% SDS for 5 min.and then placed in a 35×150 mm roller tube (052-002, Biometra, Tampa)with the DNA side facing the center of the tube. Next, 5 mlhybridization solution (HYB125.GF, Research Genetics), 5 μl Poly(dA)(POLYA.GF, Research Genetics) and 5 μl Cot-1 DNA (15279-011, LifeTechnologies) were added to the tube, that was placed in a 42° C.hybridization oven for 2 to 4 hours.

[0161] DNA for hybridization on gene filter was labeled as follows.Total RNA (0.8 μg) was suspended to 8 μl RNase-free water. Two μl of 1μg/μl 10-20 mer of Oligo-(dT) (POLYT.GF, Research Genetics) was added tothe RNA solution in a tube that was then incubated in a 70° C. for 10min. Then, the tube was briefly chilled on ice. Next, 6 μl 5× FirstStrand Buffer (18064-014, Life Technologies), 1 μl of 0.1M DTT(18064-014, Life Technologies), 1.5 μl of 100 mM dNTP (27-2035-02,Amersham Pharmacia), 1.5 μl Superscript II reverse transcriptase(18064-014, Life Technologies) and 10 μl ³³P dCTP (BF1003, AmershamPharmacia) were added and mixed thoroughly. A count per minute forradioactivity was recorded by use of Scanner QC4000 (Bioscan Inc.Washington, D.C.). The mixture was placed in a 37° C. water bath for 90min.

[0162] The labeled DNA was brought up to 100 μl Rnase-free water andthen purified by use of a Bio-Spin 6 chromatography column (732-6002,Bio-Rad, Hercules, Calif.) following the manufacturer's instruction. DNAwith more than 5% of α-³³P incorporation was denatured for 5 min in aboiling water bath and added directly to the pre-hybridization. Thehybridization was allowed to continue for 15 h at 42° C. The washes weredone to the final stringency of 0.5×SSC, 1% SDS at 50° C. for 15 min.The filters were placed on ddH₂O-moistened piece of Whatmann paper(28458-005, VWR, Bridgeport, N.J.), exposed onto a phosphor screen(Molecular Dynamics) for 5 h, and scanned for signals with the Storm 840Scanner (Molecular Dynamics). The tiff images were transferred tosoftware IPLab/ArraySuite v2.0 (NHGRI/NIH) for identification ofdifferentially expressed genes as described previously (Su et al., Mol.Carcinog., 28:119-127, 2000).

[0163] Based on selection criterions of at least 800 expressionintensities and 2-fold differences between the two cell lines, 651 of19,592 genes (3.32%) (FIG. 1, panels C and D) were selected for makingmicroarrays on glass slides to further investigate their expression inmultiple breast cancer samples.

Example 2 Customized cDNA Microarrays on Glass Slides

[0164] In order to reproducibly measure gene expression, the resultant651 differentially expressed genes and 117 controls were printed asdouble sets on the individual glass slides. The same batch microarrayswere used to measure gene expression in MDA-MB-231 and MDA/H6 celllines. Briefly, human sequence verified unigene cDNA clones werepurchased from Research Genetics. Plasmid DNAs were isolated frombacterial clones. cDNA inserts were amplified by PCR using the vectorsequence-specific primers flanking the inserts. 0.21 μg/ml of thepurified products including 651 cDNAs, 80 housekeeping genes for ratiocontrol (Chen et al. Biomed. Optics, 2:364-374, 1997), 4 non-specificcontrols of E. coli DNA, and 33 negative controls of non-DNA sample wereprinted as double sets on the individual glass slides using GMS417arrayer (Affymetrix).

[0165] The first strand cDNA was labeled by using MicroMax Kit (NEN,Boston, Mass.) following the manufacturer's instruction. All cancersamples were labeled with the fluorescent Cy3-dUTP and the referencesample (MDA/H6) with Cy5-dUTP. Very briefly, 50-ug total RNA was mixedwith Cy3-dUTP (or Cy5-dUTP) and other reagents from the kit tosynthesize the label first strand cDNA at 42° C. for h. The reaction wasstopped by addition of 2.5 ul 0.5M EDTA and 2.5 ul 1N NaOH and thenincubated at 65° C. for 30 min. After adding 6.2 ul 1M Tris-HCl (pH7.5), the samples were purified by use of Microcon 100 (Cat. No. 42412,Millipore Corp., Bedford, Mass.) to remove unincorporated nucleotidesand salts. The Cy3- and Cy5- labeled DNA samples of each pair weredissolved into 25 μl Hybridization Buffer from the kit by heating at 50°C. for 10 min. After overlaying a cover slip onto a microarrayed glassslide, the DNA sample was heated at 90° C. for 2 min. After a quickspin, 25-ul sample was placed onto the edge of the coverslip. The samplewas drawn underneath the coverslip by capillary action. Each slide wasplaced in a 50-ml conical tube with moisture Kimwipe. Hybridization wasallowed to proceed at 65° C. for 16 h. The slides were washed to a finalstringency of 0.06×SSC at room temperature for 15 min.

[0166] Image and Statistic Analysis

[0167] Hybridized array slides were scanned by use of GenePix 4000ALaser Scanner (Axon Instruments, Inc., Foster City, Calif.). For eachslide, two fluorescent intensities (Cy3, Cy5) were scanned separatelyand then placed into the red and green channel as the tiff images insoftware IPLab/ArraySuite v2.0 (NHGRI, NIH) for analysis.

[0168] Image segmentation, target detection and ratio calibrationmethods were employed to report the expression ratios of each gene onthe slides (Sorlie Proc. Natl. Acad. Sci. U.S.A, 98:10869-10874, 2001).The ratio calibration on gene filters were performed based on signalintensities of all the targets; whereas the ratio calibration on glassslides were conducted based on 80 pre-selected internal control genes ofwhich ratios were normalized close to a value of 1.0. A 99% confidenceinterval was used to determine significantly up- and down-expressedgenes. In addition, an empirically determined intensity filter (greaterthan 800 on gene filters or greater than 2,000 of average intensities inred or green channels on glass slides, for an intensity range from 0 to65,535) was applied to further strengthen the stringency for analysis.Scatter plots were drawn in which the calibrated ratios of genes fromone set were plotted against those of the other on a log-scale. Thelinear regression and Pearson coefficient of correlation computed fromthe scatter plots were used to interpret the strength of the relationsof gene expression detected by two sets of genes on the same slides andby genes on two different slides. Multidimensional scaling analysis wasperformed by use of software developed under MatLab 5.2.1 (TheMathWorks, Inc.) platform for the Mac computer. Hierarchical dendrogramclustering analysis was conducted by using the software Cluster/TreeView(Eisen et al. Proc. Natl. Acad. Sci. U.S.A, 95:14863-14868, 1998).

[0169] Panels A and B of FIG. 2 show the representative image of 2 setsof genes on the same slide. The calibrated expression ratios ofinformative genes (>2,000 average intensities in either red or greenchannel) from these two cell lines were subjected to log-transformationto obtain approximate normal distribution. The log-transformed ratiosfrom one set of genes were drawn against those from the other as ascatter plot, from which a linear regression and Pearson coefficient ofcorrelation were computed. Panels C and D of FIG. 2 show the strongpositive linear relations between Set A and Set B on Slide 1 and Slide2, respectively. In addition, Pearson coefficient of correlation betweenthe Set A and the Set B on Slide 1 and Slide 2 were 0.986 and 0.974,respectively. The expression ratios of genes from Set A and Set B wereaveraged for the same slides. The average values from Slide 1 wereplotted against those from Slide 2. The results indicated, again, astrong positive linear relation with the high value of Pearsoncoefficient of correlation (r=0.982) (Panel E, FIG. 2), demonstratingthe strength of reproducibility of the slides and the experiments.

Example 3 Gene Expression Profile of 13 Breast Cancer Samples

[0170] The high quality cDNA microarrays were used to measure expressionof 768 arrayed elements (651 differentially expressed genes and 117controls) in 13 malignant breast cancers using the non-tumorigenic cellline MDA/H6 as a common reference. RNA samples were purified from breastcancer cell lines (n=10) and breast cancer tissues (n=3) (Table 3) andlabeled by Cy3-dUTP for microarray hybridization. The reference MDA/H6samples were labeled with the Cy5-dUTP. An additional MDA-MB-231 sampleand a melanoma sample were used as controls for identity anddissimilarity, respectively. Thus, a total of 15 experiments wereperformed. Out of 731 arrayed human genes, 202 (27.63%) passed thescreening filter of the average intensities of genes in red or greenchannel greater than 2,000 in the range from 0 to 65,535. The expressionratios of the 202 genes were used to compute Pearson coefficient ofcorrelation (or similarities and dissimilarities) among the samples andamong the genes. The relative relations of these cancer samples werevisualized by multidimensional scaling analysis (MDS, Panel A, FIG. 3)and hierarchical clustering analysis (Panel C, FIG. 3). Panel B of FIG.3 shows the gene dendrogram from the hierarchical clustering analysis.These results revealed that, first, the expression profiles of twoMDA-MB-231 samples were essentially identical (r=0.9823) and that,secondly, the expression pattern of the melanoma sample was the mostdissimilar to that of the MDA-MB-231 (r=0.325), as expected. Thirdly,the expression patterns of all other breast cancer samples weredistributed between the identical and dissimilar controls (MDA-MB-231and melanoma). Finally, Pearson coefficients of correlation betweenbreast cancer cell lines BT20, BT474 and ZR75-1 were 0.796, indicatingtheir similarities.

Example 4 Frequently Differentially-Expressed Genes

[0171] Microarray gene expression analysis revealed 19 genes with highfrequent alterations in their expression in human breast cancers. Out of202 genes with informative expression levels, 9 were highlyover-expressed (Panel D, FIG. 3) and 10 were significantlydown-regulated (Panel E, FIG. 3) in at least 10 of 13 breast cancersamples. Twenty-one had no significant changes in expression in all 13breast cancer samples and the remaining 162 genes displayed more than 2fold changes in at least 1 of 13 samples studied. The nine up-regulatedgenes are listed in Table 4. The ten down-regulated genes are listed inTable 5.

Example 5 The Decrease of the THBD Protein in Breast Cancer Cell Linesand Tissue Specimens

[0172] The microarray analysis showed a range from 3 fold to more than10 fold down-regulation of the THBD RNA in all 13 human breast cancersstudied (Panel A, FIG. 4). In order to determine the levels of the THBDprotein, Western blot analysis was performed on the breast cancer celllines MDA/H6, MDA-MB-231, MDA-MB-436, MDA-MB-453, and BT549 (Panel B,FIG. 4). Briefly, cells at 80% confluenc were rinsed twice with ice-coldPBS, scraped into a microcentrifuge tube and pelleted by centrifugationat 6,000 rpm at 4° C. for 3 min. The cell pellets were resuspended in500 μl Lysis Buffer (1% NP40, 1% sodium deoxycholate, 0.1% SDS, 150 mMNaCl, 0.01M Na₂HPO₄, pH7.4, 1 μg/ml proteinase inhibitors). The lysatewere spun at 14,000 rpm at 4° C. for 5 min, after which the supernatantswere transferred to a fresh ice-chilled microcentrifuge tube. Proteinwas then assayed using the Pierce BCA Protein Assay kit (Microwell PlateProtocol) (Pierce, Cat# 23225, Rockford, Ill.). For each sample, theprotein concentration was adjusted to 10 μg/μl. Five μl of each samplewas mixed with equal volume of 2× loading dye (SeeBlue Pre-StainedStandard, Cat# LC5625, Invitrogen), heated for 5 min at 95° C. and thenloaded onto the 8% SDS-polyacrylamide gel (Cat# EC6045, Invitrogen) inthe Minigel apparatus (XCELLII, Cat# EI9051, Invitrogen). The gel wasrun at 150V for 1-1.5 h. The proteins were transferred from the gels tonitrocellulose membrane by use of blotting pads (XCELLII Blotting, Cat#EI9052, Invitrogen) for 1 h under 30V. The membranes were submerged inblocking solution (2.5 g non fat dry milk, 47.5 ml 1× TBS and 20 μlTween 20) for 1 h at room temperature. The membrane was then rinsed withthe blocking solution, and then incubated in the solution of polyclonalgoat antibody of thrombomodulin (1:200 dilution with the blockingsolution) (Cat# SC-7096, Santa Cruz Biotechnology, Santa Cruz, Calif.)for 1 h at room temperature. The primary antibody was rinsed off withwashing solution (49.95 ml 1× TBS and 25 μl Tween 20) three times for 5min each. The membrane was then incubated in the solution ofanti-goat-IgG-HRP (1:1,000 dilutions) (Cat# sc-2056, Santa CruzBiotechnology) for 1 h at room temperature. The secondary antibody waswashed off with the washing solution for 3 times, 10 min each and oncewith 1× TBS for 15 min. The membrane was incubated in an enhancedchemiluminescent substrate (Pierce Supersignal ChemiluminescentSubstrate, Cat# 34080, Pierce, Rockford, Ill.) for min, wrapped in SaranWrap, and exposed to Kodak X-Omat AR film at room temperature for 2 secto 1 min. The goat polyclonal IgG of actin I-19 (Cat# sc1616, Santa CruzBiotechnology) was used as a loading control.

[0173] The results demonstrated the high level of the THBD protein innon-tumorigenic breast cancer cell line MDA/H6. In contract, it wasdecreased approximately 5 folds in MDA-MB-231 and 3 folds in MDA-MB-453,and was not detectable in MDA-MB-436 and BT549. Thus, the resultscorrelated the THBD RNA levels to the protein expression, that is, bothof the RNA and the protein were decreased in the breast cancer samples.

[0174] In situ immunohistochemical staining for THBD protein wasconducted on 20 cases of breast normal and cancer tissue specimens inorder to determine THBD protein levels in vivo. Briefly, the tissuesections on slides were incubated at a 60° C. for 1 h, and then immersedin Xylenes (X5-500, Fisher Healthcare, Hanover Park, Ill.) at roomtemperature for 5 min, twice. The slides were re-hydrated by immersingconsecutively in 100%, 75% and 50% ethanol alcohol at room temperature,2 min in each solution and twice per solution. The slides were rinsedwith ddH₂O for 5 min and then immersed into 10 mM Sodium Acetate buffer(pH: 6.0) in a plastic box that was incubated in boiling water for 10min. All the following procedures were carried out at room temperature.The slides were rinsed with 1× Phosphate Buffered Saline (PBS) (FisherHealthcare, Hanover Park, Ill.) for 5 min, and then incubated in 3%peroxide (Fisher Healthcare, Hanover Park, Ill.) for 10 min. Afterwashed with 1× PBS buffer for 3 min, twice, the slides were mounted onShandon chamber coverslip (Shandon Inc, Pittsburgh, Pa.). From now on,the slides were washed with Cadenza Buffer (407340, Shandon, Inc.) for 4min, referring as washing in the following procedures. Two hundred μl ofProtein Block (HK112-9K, BioGenex, Inc.) was placed onto each slide,incubating for 20 min. TM(C-17), an affinity purified goat polyclonalantibody against a peptide at the carboxyl terminus of humanthrombomodulin (Santa Cruz, Inc.), was diluted with 1% BSA and 0.01%NaAzide solution to 200- 400 folds. After washing the slides, 200 μl ofthe diluted antibody was dropped onto each slide, incubating for 1 h.Then, the sections were processed in the following order: incubation in200 μl anti-immunoglobulin (HK340-9K, BioGenex, Inc.) for 20 min,washing, incubation in 200 μl peroxidase-conjugated streptavidin(HK330-9K, BioGenex, Inc.) for 20 min, washing, incubation in 200 μl DAB(3,3′-diaminobenzidine) Chromogen (HK153-5K, BioGenex, Inc.) for 10 min,and washing. Each slide was counterstained with 300 μl of hematoxylin(HK100-9K, BioGenex, Inc.) for 4 min and then rinsed with ddH₂O for 3min. The sections were dehydrated by immersing consecutively in 50%,75%, and 100% ethanol alcohol for 1 min, twice in each solution. Afterrinsing in Xylenes for min, twice, the slides were mounted forvisualization under microscope. Negative controls were processed in thesame procedures as above in the absence of the antibody TM(C-17).

[0175] The in situ immunohistochemical staining demonstrated strongpositive THBD stain in normal mammary epithelial cells and negative inbreast cancer cells (FIG. 5). The control staining for both normal andbreast cancer sections without the antibody were negative. Table 6summarizes the results that 18 out of the 20 cases, including all 5metastatic breast cancer samples and 13 infiltrating ductal carcinomasamples, lost the THBD protein in the cancer cells, and one case ofmoderately well differentiated infiltrating adenocarcinoma and one caseof infiltrating ductal carcinoma with intramammary lymphatic invasionhad the cancer cells with the positive THBD stain. Thus, the resultsindicated that the THBD protein were absent in advanced breast cancers.

[0176] From the foregoing it will be appreciated that, although specificembodiments of the invention have been described herein for purposes ofillustration, various modifications may be made without deviating fromthe spirit and scope of the invention. Accordingly, the invention is notlimited except as by the appended claims.

1 38 1 2787 DNA Homo sapiens 1 agagcggagg ccgcactcca gcactgcgcagggaccgcct tggaccgcag ttgccggcca 60 ggaatcccag tgtcacggtg gacacgcctccctcgcgccc ttgccgccca cctgctcacc 120 cagctcaggg gctttggaat tctgtggccacactgcgagg agatcggttc tgggtcggag 180 gctacaggaa gactcccact ccctgaaatctggagtgaag aacgccgcca tccagccacc 240 attccaagga ggtgcaggag aacagctctgtgataccatt taacttgttg acattacttt 300 tatttgaagg aacgtatatt agagcttactttgcaaagaa ggaagatggt tgtttccgaa 360 gtggacatcg caaaagctga tccagctgctgcatcccacc ctctattact gaatggagat 420 gctactgtgg cccagaaaaa tccaggctcggtggctgaga acaacctgtg cagccagtat 480 gaggagaagg tgcgcccctg catcgacctcattgactccc tgcgggctct aggtgtggag 540 caggacctgg ccctgccagc catcgccgtcatcggggacc agagctcggg caagagctcc 600 gtgttggagg cactgtcagg agttgcccttcccagaggca gcgggatcgt gaccagatgc 660 ccgctggtgc tgaaactgaa gaaacttgtgaacgaagata agtggagagg caaggtcagt 720 taccaggact acgagattga gatttcggatgcttcagagg tagaaaagga aattaataaa 780 gcccagaatg ccatcgccgg ggaaggaatgggaatcagtc atgagctaat caccctggag 840 atcagctccc gagatgtccc ggatctgactctaatagacc ttcctggcat aaccagagtg 900 gctgtgggca atcagcctgc tgacattgggtataagatca agacactcat caagaagtac 960 atccagaggc aggagacaat cagcctggtggtggtcccca gtaatgtgga catcgccacc 1020 acagaggctc tcagcatggc ccaggaggtggaccccgagg gagacaggac catcggaatc 1080 ttgacgaagc ctgatctggt ggacaaaggaactgaagaca aggttgtgga cgtggtgcgg 1140 aacctcgtgt tccacctgaa gaagggttacatgattgtca agtgccgggg ccagcaggag 1200 atccaggacc agctgagcct gtccgaagccctgcagagag agaagatctt ctttgagaac 1260 cacccatatt tcagggatct gctggaggaaggaaaggcca cggttccctg cctggcagaa 1320 aaacttacca gcgagctcat cacacatatctgtaaatctc tgcccctgtt agaaaatcaa 1380 atcaaggaga ctcaccagag aataacagaggagctacaaa agtatggtgt cgacataccg 1440 gaagacgaaa atgaaaaaat gttcttcctgatagataaaa ttaatgcctt taatcaggac 1500 atcactgctc tcatgcaagg agaggaaactgtaggggagg aagacattcg gctgtttacc 1560 agactccgac acgagttcca caaatggagtacaataattg aaaacaattt tcaagaaggc 1620 cataaaattt tgagtagaaa aatccagaaatttgaaaatc agtatcgtgg tagagagctg 1680 ccaggctttg tgaattacag gacatttgagacaatcgtga aacagcaaat caaggcactg 1740 gaagagccgg ctgtggatat gctacacaccgtgacggata tggtccggct tgctttcaca 1800 gatgtttcga taaaaaattt tgaagagttttttaacctcc acagaaccgc caagtccaaa 1860 attgaagaca ttagagcaga acaagagagagaaggtgaga agctgatccg cctccacttc 1920 cagatggaac agattgtcta ctgccaggaccaggtataca ggggtgcatt gcagaaggtc 1980 agagagaagg agctggaaga agaaaagaagaagaaatcct gggattttgg ggctttccag 2040 tccagctcgg caacagactc ttccatggaggagatctttc agcacctgat ggcctatcac 2100 caggaggcca gcaagcgcat ctccagccacatccctttga tcatccagtt cttcatgctc 2160 cagacgtacg gccagcagct tcagaaggccatgctgcagc tcctgcagga caaggacacc 2220 tacagctggc tcctgaagga gcggagcgacaccagcgaca agcggaagtt cctgaaggag 2280 cggcttgcac ggctgacgca ggctcggcgccggcttgccc agttccccgg ttaaccacac 2340 tctgtccagc cccgtagacg tgcacgcacactgtctgccc ccgttcccgg gtagccactg 2400 gactgacgac ttgagtgctc agtagtcagactggatagtc cgtctctgct tatccgttag 2460 ccgtggtgat ttagcaggaa gctgtgagagcagtttggtt tctagcatga agacagagcc 2520 ccaccctcag atgcacatga gctggcgggattgaaggatg ctgtcttcgt actgggaaag 2580 ggattttcag ccctcagaat cgctccaccttgcagctctc cccttctctg tattcctaga 2640 aactgacaca tgctgaacat cacagcttatttcctcattt ttataatgtc ccttcacaaa 2700 cccagtgttt taggagcatg agtgccgtgtgtgtgcgtcc tgtcggagcc ctgtctcctc 2760 tctctgtaat aaactcattt ctagcag 27872 424 DNA Homo sapiens 2 accagcccag cctttcagtg caggctccag ccctccacccccacccgagt tgcaggatgt 60 cgatgacaga cttgctgaac gctgaggaca tcaagaaggcggtgggagcc tttagcgcta 120 ccgactcctt cgaccacaaa aagttcttcc aaatggtcggcctgaagaaa aagagtgcgg 180 atgatgtgaa gaaggtgttt cacatgctgg acaaggacaaaagtggcttc atcgaggagg 240 atgagctggg attcatccta aaaggcttct ccccagatgccagagacctg tctgctaaag 300 aaaccaagat gctgatggct gctggagaca aagatggggacggcaaaatt ggggttgacg 360 aattctccac tctggtggct gaaagctaag aagcactgactgcccctggt cttccacctc 420 tctg 424 3 6455 DNA Homo sapiens 3 gaggaagcgattctggggtt tctgtgttga acggttcttg tccgcgaaga tgcgcttcgg 60 cctctgtcaggggacttgaa cgggcttagt gggcttcagc cagcttttct ccaccggttc 120 cccacggggacccccccccc ccggccgttg caatggcggg cgtggggccg gggggctacg 180 cggcggagttcgtgccaccg ccagagtgcc ccgtctttga gccgagttgg gaggagttca 240 cagatccgctcagctttatc ggccgcatcc ggcctttggc ggagaaaacc ggcatctgca 300 aaattcggccgcccaaggac tggcagcctc catttgcctg tgaagtaaaa agctttcgtt 360 tcactccaagagtccagcgc ctgaatgaac ttgaggcaat gaccagagtg agattggatt 420 tcttggatcaactagcaaaa ttttgggaac ttcaaggatc tactctgaag atccctgtgg 480 tagagagaaaaatcctggat ctgtatgctt tgagcaagat tgttgccagc aaaggaggtt 540 ttgaaatggtcaccaaagag aagaaatggt ctaaagtggg tagtcgcttg ggatatctgc 600 caggaaaaggaactgggtct cttttgaagt cacattatga aagaattctc tacccatatg 660 agcttttccagtctggtgtg agccttatgg gtgtgcagat gcctaattta gatcttaaag 720 aaaaagtggagcctgaggtt ctcagcactg atacccaaac ttccccagag ccaggcacaa 780 ggatgaacattctgccgaag agaacaagac gtgtgaagac tcagtcagaa tctggagatg 840 tgagtagaaacacggaactg aagaaacttc agatttttgg ggctgggccc aaggttgtgg 900 gcttggcaatgggaacaaaa gataaagaag atgaggtcac ccgaagacga aaagttacca 960 acaggtcagacgcatttaac atgcaaatga gacaacggaa aggcactctc tctgttaact 1020 ttgttgatctctatgtttgt atgttttgtg gtcggggaaa caatgaagat aaattgcttt 1080 tgtgtgatggatgtgatgac agctatcata cattttgtct aattcctcca ctacctgatg 1140 tgcccaaaggagactggagg tgtcctaaat gtgtcgccga ggaatgtagc aaacctcgag 1200 aagcctttggatttgaacaa gctgtacgag agtatacact tcagagcttt ggagagatgg 1260 cagataattttaagtctgat tattttaata tgccagtcca tatggttccc acagaactag 1320 tagaaaaggaattttggcgg ctggtaagca gcattgaaga agatgttatt gtggaatatg 1380 gagcagatatctcctcaaaa gactttggaa gtggatttcc ggtgaaggat gggcggagaa 1440 agattctgccagaagaagag gaatatgcac tttctggttg gaatttgaat aacatgcctg 1500 tcctggaacagtctgttctt gcacatatta atgtggacat ctctggtatg aaagtgccgt 1560 ggctctatgtgggaatgtgc ttctcttctt tttgctggca cattgaggat cactggagtt 1620 attccatcaactacttgcac tggggggagc caaagacatg gtatggtgtg ccatctcatg 1680 ctgcagagcaactggaggag gtgatgagag agctggcccc cgagttattt gaatcccagc 1740 ctgatcttctgcatcagtta gttaccatca tgaaccccaa cgtgctaatg gagcatggtg 1800 tgcctgtgtacaggaccaat cagtgtgctg gcgagtttgt tgtgacattt cctcgtgcct 1860 atcactctggatttaaccag ggctacaact ttgctgaagc tgtgaacttc tgtactgctg 1920 actggttgcccattggacgt caatgtgtaa atcattaccg acgcctaagg cgccactgtg 1980 tcttttcacacgaggaacta attttcaaga tggcagcaga tccagaatgc ttagatgtgg 2040 ggctggctgccatggtctgc aaagaattga ctctcatgac tgaagaagaa acacgattaa 2100 gagagtctgttgtacagatg ggtgtcctga tgtcagaaga agaagtgttt gaacttgttc 2160 ctgatgatgagcggcagtgt tcagcatgca gaaccacatg ttttctctct gctctcacat 2220 gttcctgtaatcctgagcgg cttgtatgtc tctaccatcc aactgatctg tgcccctgcc 2280 ccatgcagaagaaatgtctt agatatcgct acccattaga agacctccct tctctgctat 2340 atggtgtaaaagtcagggca cagtcctatg acacttgggt cagtcgtgtt acagaagcat 2400 tgtctgctaacttcaaccac aaaaaagatt tgattgaatt gcgagtaatg ctggaagatg 2460 ctgaggataggaaataccca gagaatgatc tctttcgaaa actcagggat gctgtaaaag 2520 aagctgagacctgtgcttct gtggctcagc tgcttctgag caaaaagcag aaacacagac 2580 agagcccagatagtgggagg actcggacca aactgacagt ggaagaattg aaggcctttg 2640 tccaacaactttttagtctt ccgtgtgtca tcagccaagc tcggcaagta aagaatctgc 2700 tagatgatgtggaagagttt catgaacgtg ctcaggaggc catgatggat gaaaccccag 2760 attcttccaaactccagatg ttgatagata tgggctctag tctctatgtg gaactccctg 2820 aattaccacgactgaagcaa gagctacaac aggctcggtg gttggacgaa gtaagactga 2880 ccttatcagatccgcaacaa gtcactttgg atgtcatgaa gaagctgata gactctgggg 2940 tagggttggcaccccaccat gctgtggaga aagcaatggc tgaactacag gagctcctta 3000 cagtctctgaacgatgggaa gaaaaggcta aggtctgcct acaggcaaga ccgaggcaca 3060 gtgtggcaagtttagaaagc attgtgaatg aagccaagaa cattccagcc tttctaccca 3120 atgtgttgtccttgaaagaa gccttacaaa aggctcgaga atggaccgct aaagtggaag 3180 ctattcagagtggcagcaat tacgcttatt tggagcagct tgagagcttg tctgcgaaag 3240 gacgccctattcctgtgcgt cttgaagcac tgccgcaagt ggaatcacag gtagcagcag 3300 cacgggcatggagagaacgg actgggcgga cgtttcttaa gaagaattct agccatacat 3360 tgttacaggtgctgagcccc cggaccgaca ttggtgtata tgggagtggc aaaaatagga 3420 ggaaaaaagtaaaagaacta atagaaaaag aaaaagaaaa ggatctggac ctggagcctc 3480 tgagtgatctggaggaagga ttggaggaaa ccagagatac agccatggtg gtggcagttt 3540 tcaaagaacgggagcaaaaa gagattgaag ccatgcattc tctcagagca gccaacctag 3600 ccaagatgacaatggtggac cgcatagaag aagtaaaatt ttgcatttgc cgcaagacag 3660 ccagtgggtttatgctacag tgtgagctct gcaaagactg gttccataac agctgtgttc 3720 ctcttcctaaatcaagttcc caaaaaaaag gatccagctg gcaagctaaa gaagtaaaat 3780 tcctttgccctctttgtatg cggtctcgaa ggcccaggct agagactatt ctgtcactcc 3840 tggtatcccttcagaagttg cccgtacggt tgcctgaagg agaggccctg cagtgtttga 3900 cagaacgtgctatgagttgg caagatagag cgcggcaggc tctagccaca gatgaactat 3960 cctctgccctggccaaacta tctgtgttga gccagcgtat ggtggaacag gcggctcgag 4020 aaaaaactgaaaagatcatc agtgcagaac tccaaaaagc agctgccaat ccagacttac 4080 agggacacttacctagtttc cagcagtctg cttttaaccg ggtggtgagc agtgtgtcat 4140 cttctcctcgacaaacaatg gactatgatg atgaagaaac agactctgat gaagacattc 4200 gagagacatatggctacgac atgaaggaca cagccagtgt gaagtcctct agtagtcttg 4260 aacccaatcttttttgtgat gaagagattc ccatcaaatc cgaggaggtg gtgacccaca 4320 tgtggacagcaccttcattt tgtgcagagc atgcttattc ttctgcttct aagagttgtt 4380 ctcaagtattttttgggaaa ggttctagca ccccaaggaa acaacctcgg aagagccctt 4440 tggtgccccgaagtttggaa cctccagtgc tggagttgtc acctggagct aaggcacaac 4500 tggaagaacttatgatggtt ggagatctcc tggaagtatc tctggacgag actcaacaca 4560 tatggcggattttgcaggcc acacacccac cctctgaaga cagattcttg catatcatgg 4620 aggatgacagcatggaagag aaaccactaa aagtgaaagg aaaggactct tcagagaaga 4680 aacggaaacggaagctagaa aaggtagagc aactttttgg agaaggaaaa cagaagtcca 4740 aggagttaaagaaaatggac aaacctagaa agaagaaatt aaaattaggt gcagacaaat 4800 caaagaagctgaataaactg gccaagaaac tagcaaaaga agaagagaga aagaaaaaga 4860 aggagaaggctgctgcagcc aaagttgaac ttgtgaaaga gagcactgaa aagaaaagag 4920 agaaaaaggtgctggacatc ccctcaaagt atgactggtc aggagcagag gagtctgatg 4980 atgagaatgctgtgtgcgca gaaccagact gccaaaggcc ctgcaaggac aagggagttg 5040 tatttgtaacgaagaagaga gagataaaaa atattagttt taaaagtgtc ctatgtgact 5100 gcttttctaaaaaggtagac tgggtacaat gtgatggtgg ctgtgatgag tggtttcatc 5160 gggtttgtgtgggtgtatct ccagaaatgg ctgaaaatga agattacatc tgtataaact 5220 gtgcaaagaagcaggggcca gttagcccag gtccagcacc acctccttcc ttcataatga 5280 gctacaaactaccaatggag gatcttaaag agaccagtta gcagatgctt ggttagtttg 5340 ggacatggggggacatggac cacattgaga ccttagtcat caagtagagt ggtttatatc 5400 acttggaatgttgcttctaa agatgaatgg ccttcagaga aagtcccctt agtgctggct 5460 tcctctttgcatggactctg tgggttacat tgctctatca acatatctat gcagagggtg 5520 tcttctttggtacaacagcc aatatctcat gtctcctttg agtgtggttt actgcattaa 5580 ggccagatgcttaattgagc tctagggtgg ctggttagta ttaatacatt ggtgtgctaa 5640 cagggcatataggatgtggc ttttgtccag ctgatagtag ttagaggctt acaacttagg 5700 agcagcaccaactgaaggtg ctaattgctt ggatctcctt cattaggata gttggagagg 5760 gattggagtaccactttctt ccactgttac caggtactta atgccctaaa gatacaacta 5820 ggagtaacagggccaaagtt atttctgtta gacgtcaagg aatggtatca cagtctattg 5880 acctcagcgatttgtgcttg tttgtgctag aagaacatcc caaataggag aacctctcac 5940 aagctggggcaggtcacctt atctttgtaa gatgaggata tcatctagat cagaaatctg 6000 actagattggattctgagga gaagaaccta ctacaaggca aggagccgtt ttttggcttt 6060 gaaaagtcttgctgtcttgg gtctacattt tagggaagag caggtacatg gatccaggct 6120 tctgccaaaaaaaaaaagag aagaagatga cgagtatgac cagtcgtact atcttactga 6180 gccacagtgatgcatgcttt tcggggaaaa cttcattcac aagtattcca gacaccaggc 6240 ttcaggcatggccatgagca agaccagcaa ataacagctt tttcccttgc agccctgacc 6300 ccaatgtctgctgtttccaa cactggtgat ttctaactac ggcccacagc agatgctgtt 6360 gaataacaccatggcttcat cagaggatgt ggggttgtag tacctctggg tgatgaagtt 6420 gttttagcaaatccattttt aaaaaaaaaa aaaaa 6455 4 1369 DNA Homo sapiens 4 ggccgacagtgcctgatttg agatggggtc ccaggtctcg gtggaatcgg gagctctgca 60 cgtggtgattgtgggtgggg gctttggcgg gatcgcagca gccagccagc tgcaggccct 120 gaacgtccccttcatgctgg tggacatgaa ggactccttc caccacaatg tggctgctct 180 ccgagcctccgtggagacag ggttcgccaa aaagacattc atttcttact cggtgacttt 240 caaggacaacttccggcagg ggctagtagt ggggatagac ctgaagaacc agatggtgct 300 gctgcagggtggcgaggccc tgcccttctc tcatcttatc ctggccacgg gcagcactgg 360 gcccttcccgggcaagttta atgaggtttc cagccagcag gccgctatcc aggcctatga 420 ggacatggtgaggcaggtcc agcgctcacg gttcatcgtg gtggtgggag gaggctcggc 480 tggagtggagatggcagcag agattaaaac agaatatcct gagaaagagg tcactctcat 540 tcactcccaagtggccctgg ctgacaagga gctcctgccc tccgtccggc aggaagtgaa 600 ggagatcctcctccggaagg gcgtgcagct gctgctgagt gagcgggtga gcaatctgga 660 ggagctgcctctcaatgagt atcgagagta catcaaagtg cagacggaca aaggcacaga 720 ggtggccaccaacctggtga ttctctgcac cggcatcaag atcaacagct ccgcctaccg 780 caaagcatttgagagcagac tagccagcag tggtgctctg agagtgaacg agcacctcca 840 ggtggagggccacagcaacg tctacgccat tggtgactgt gccgacgtga ggacgcccaa 900 gatggcctatcttgccggcc tccacgccaa catcgccgtg gccaacatcg tcaactctgt 960 gaagcagcggcctctccagg cctacaagcc gggtgcactg acgttcctcc tgtccatggg 1020 gagaaatgacggtgtgggcc aaatcagtgg cttctatgtg ggccggctca tggttcggct 1080 gaccaagagccgggacctgt tcgtctctac gagctggaaa accatgaggc agtctccacc 1140 ttgatggagaggccaggcgg gagaactacc gcagcaggtg ggcgtacgga ctgcttggcg 1200 catggcacccgcctggcaag tgctagaact aatgctattc ttctggaata agatgccaat 1260 gatgtggtggctagaaatgc aacttgtata aaacaaaaat gggagagaga gaggtattaa 1320 acaaataccccccttagagg ataaaaaaaa aaaaaaaaaa aaaaaaaaa 1369 5 1712 DNA Homo sapiens5 ggcacgaggg gcagctgtcg gctggaagga actggtctgc tcacacttgc tggcttgcgc 60atcaggactg gctttatctc ctgactcacg gtgcaaaggt gcactctgcg aacgttaagt 120ccgtccccag cgcttggaat cctacggccc ccacagccgg atcccctcag ccttccaggt 180cctcaactcc cgcggacgct gaacaatggc ctccatgggg ctacaggtaa tgggcatcgc 240gctggccgtc ctgggctggc tggccgtcat gctgtgctgc gcgctgccca tgtggcgcgt 300gacggccttc atcggcagca acattgtcac ctcgcagacc atctgggagg gcctatggat 360gaactgcgtg gtgcagagca ccggccagat gcagtgcaag gtgtacgact cgctgctggc 420actgccgcag gacctgcagg cggcccgcgc cctcgtcatc atcagcatca tcgtggctgc 480tctgggcgtg ctgctgtccg tggtgggggg caagtgtacc aactgcctgg aggatgaaag 540cgccaaggcc aagaccatga tcgtggcggg cgtggtgttc ctgttggccg gccttatggt 600gatagtgccg gtgtcctgga cggcccacaa catcatccaa gacttctaca atccgctggt 660ggcctccggg cagaagcggg agatgggtgc ctcgctctac gtcggctggg ccgcctccgg 720cctgctgctc cttggcgggg ggctgctttg ctgcaactgt ccaccccgca cagacaagcc 780ttactccgcc aagtattctg ctgcccgctc tgctgctgcc agcaactacg tgtaaggtgc 840cacggctcca ctctgttcct ctctgctttg ttcttccctg gactgagctc agcgcaggct 900gtgaccccag gagggccctg ccacgggcca ctggctgctg gggactgggg actgggcaga 960gactgagcca ggcaggaagg cagcagcctt cagcctctct ggcccactcg gacaacttcc 1020caaggccgcc tcctgctagc aagaacagag tccaccctcc tctggatatt ggggagggac 1080ggaagtgaca gggtgtggtg gtggagtggg gagctggctt ctgctggcca ggatggctta 1140accctgactt tgggatctgc ctgcatcggt gttggccact gtccccattt acattttccc 1200cactctgtct gcctgcatct cctctgttgc gggtaggcct tgatatcacc tctgggactg 1260tgccttgctc accgaaaccc gcgcccagga gtatggctga ggccttgccc acccacctgc 1320ctgggaagtg cagagtggat ggacgggttt agaggggagg ggcgaaggtg ctgtaaacag 1380gtttgggcag tggtggggga gggggccaga gaggcggctc aggttgccca gctctgtggc 1440ctcaggactc tctgcctcac ccgcttcagc ccagggcccc tggagactga tcccctctga 1500gtcctctgcc ccttccaagg acactaatga gcctgggagg gtggcaggga ggaggggaca 1560gcttcaccct tggaagtcct ggggtttttc ctcttccttc tttgtggttt ctgttttgta 1620atttaagaag agctattcat cactgtaatt attattattt tctacaataa atgggacctg 1680tgcacaggaa aaaaaaaaaa aaaaaaaaaa aa 1712 6 2163 DNA Homo sapiens 6ggcagatgaa atataagatt catcaaccac atttgacagc ccatggcagg tttcctgttt 60tccatcgtcc ctctgcaggt cacagacaca cagagcccag ccgtggcagg ctcagccggg 120gtccggggct gctaacaacg gctacattcc tcccccaggg ccaagggaaa tcctgagcgc 180aggccagggt tgtttggttt tgaggtgtgc tgggatgaaa ggcaccctgg aagtggaagg 240taaatgaaca atggaaaaac ttcacggcaa gattagaaag atacctgagc ccaatacccg 300cctgatgtcg tgggccacac ctccgggtta ccaggggaag ggaggaagca aactgtcata 360ttgatgtggc tctaaacaac aacagtgtgc gaaggcccag gggcactttg ggattgacca 420agaggaaaca caagttgcac aatgatacaa tcttgttggt acaattgtca gagaagggaa 480ctcccacagc aaaggccata aaaccatcca gggcagtctg gggcggctca gttctgcggt 540gccagggagt ggagcagagc tcagccccgt cccaaacaca gatgggacca tgaactccgg 600acacagcttc agccagaccc cctcggcctc cttccatggc gccggaggtg gctggggccg 660gcccaggagc ttccccaggg ctcccaccgt ccatggcggt gcggggggag cccgcatctc 720cctgtccttc accacgcgga gctgcccacc ccctggaggg tcttggggtt ctggaagaag 780cagcccccta ctaggcggaa atgggaaggc caccatgcag aatctcaacg accgcctggc 840ctcctacctg gagaaggttc gcgccctgga ggaggccaac atgaagctgg aaagccgcat 900cctgaaatgg caccagcaga gagatcctgg cagtaagaaa gattattccc agtatgagga 960aaacatcaca cacctgcagg agcagatagt ggatggtaag atgaccaatg ctcagattat 1020tcttctcatt gacaatgcca ggatggcagt ggatgacttc aacctcaagt atgaaaatga 1080acactccttt aagaaagact tggaaattga agtcgagggc ctccgaagga ccttagacaa 1140cctgaccatt gtcacaacag acctagaaca ggaggtggaa ggaatgagga aagagctcat 1200tctcatgaag aagcaccatg agcaggaaat ggagaagcat catgtgccaa gtgacttcaa 1260tgtcaatgtg aaggtggata caggtcccag ggaagatctg attaaggtcc tggaggatat 1320gagacaagaa tatgagctta taataaagaa gaagcatcga gacttggaca cttggtataa 1380agaacagtct gcagccatgt cccaggaggc agccagtcca gccactgtgc agagcagaca 1440aggtgacatc cacgaactga agcgcacatt ccaggccctg gagattgacc tgcagacaca 1500gtacagcacg aaatctgctt tggaaaacat gttatccgag acccagtctc ggtactcctg 1560caagctccag gacatgcaag agatcatctc ccactatgag gaggaactga cgcagctacg 1620ccatgaactg gagcggcaga acaatgaata ccaagtgctg ctgggcatca aaacccacct 1680ggagaaggaa atcaccacgt accgacggct cctggaggga gagagtgaag ggacacggga 1740agaatcaaag tcgagcatga aagtgtctgc aactccaaag atcaaggcca taacccagga 1800gaccatcaac ggaagattag ttctttgtca agtgaatgaa atccaaaagc acgcatgaga 1860ccaatgaaag tttccgcctg ttgtaaaatc tattttcccc caaggaaagt ccttgcacag 1920acaccagtga gtgagttcta aaagataccc ttggaattat cagactcaga aacttttatt 1980ttttttttct gtaacagtct caccagactt ctcataatgc tcttaatata ttgcactttt 2040ctaatcaaag tgcgagttta tgagggtaaa gctctacttt cctactgcag ccttcagatt 2100ctcatcattt tgcatctatt ttgtagccaa taaaactccg cactagcaaa aaaaaaaaaa 2160aaa 2163 7 2856 DNA Homo sapiens 7 gtaaccgcta ctcccggaca ccagaccaccgccttccgta cacaggggcc cgcatcccac 60 cctcccggac ctaagagcct gggtcccctgtttccggagg tccgcttccc ggcccccaga 120 ttctggcatc ccagccctca gtgtccaagacccaggcagc ccgggtcccc gcctcccgga 180 tccaggcgtc cgggatctgc gccaccagaacctagcctcc tgcagacctc cgccatctgg 240 gggcactcaa cctcctggag ccaagggccccacgtcccac ccagagaaac tctcgtattc 300 ccagctccta gggccaagga acccgggcgctccgaactcc cagctttcgg acatctggca 360 cacggggcag agcagagaag ctcagcgcccagcctgggga atttaaacac tccagcttcc 420 aagagccaag gaacttcagt gctgtgaactcacaactcta aggagccctc caaagttcca 480 gtctccaggt gctgttactc aactcagtcctaggaacgtc gggtcctggg aaggagccca 540 agcgctccca gccagcttcc aggcgctaagaaaccccggt gcttcccatc atggtggccg 600 atcctcctcg agactccaag gggctcgcagcggcggagcc caccgccaac gggggcctgg 660 cgctggcctc catcgaggac caaggcgcggcagcaggcgg ctactgcggt tcccgggacc 720 aggtgcgccg ctgccttcga gccaacctgcttgtgctgct gacagtggtg gccgtggtgg 780 ccggcgtggc gctgggactg ggggtgtcgggggccggggg tgcgctggcg ttgggcccgg 840 agcgcttgag cgccttcgtc ttcccgggcgagctgctgct gcgtctgctg cggatgatca 900 tcttgccgct ggtggtgtgc agcttgatcggcggcgccgc cagcctggac cccggcgcgc 960 tcggccgtct gggcgcctgg gcgctgctctttttcctggt caccacgctg ctggcgtcgg 1020 cgctcggagt gggcttggcg ctggctctgcagccgggcgc cgcctccgcc gccatcaacg 1080 cctccgtggg agccgcgggc agtgccgaaaatgcccccag caaggaggtg ctcgattcgt 1140 tcctggatct tgcgagaaat atcttcccttccaacctggt gtcagcagcc tttcgctcat 1200 actctaccac ctatgaagag aggaatatcaccggaaccag ggtgaaggtg cccgtggggc 1260 aggaggtgga ggggatgaac atcctgggcttggtagtgtt tgccatcgtc tttggtgtgg 1320 cgctgcggaa gctggggcct gaaggggagctgcttatccg cttcttcaac tccttcaatg 1380 aggccaccat ggttctggtc tcctggatcatgtggtacgc ccctgtgggc atcatgttcc 1440 tggtggctgg caagatcgtg gagatggaggatgtgggttt actctttgcc cgccttggca 1500 agtacattct gtgctgcctg ctgggtcacgccatccatgg gctcctggta ctgcccctca 1560 tctacttcct cttcacccgc aaaaacccctaccgcttcct gtggggcatc gtgacgccgc 1620 tggccactgc ctttgggacc tcttccagttccgccacgct gccgctgatg atgaagtgcg 1680 tggaggagaa taatggcgtg gccaagcacatcagccgttt catcctgccc atcggcgcca 1740 ccgtcaacat ggacggtgcc gcgctcttccagtgcgtggc cgcagtgttc attgcacagc 1800 tcagccagca gtccttggac ttcgtaaagatcatcaccat cctggtcacg gccacagcgt 1860 ccagcgtggg ggcagcgggc atccctgctggaggtgtcct cactctggcc atcatcctcg 1920 aagcagtcaa cctcccggtc gaccatatctccttgatcct ggctgtggac tggctagtcg 1980 accggtcctg taccgtcctc aatgtagaaggtgacgctct gggggcagga ctcctccaaa 2040 attatgtgga ccgtacggag tcgagaagcacagagcctga gttgatacaa gtgaagagtg 2100 agctgcccct ggatccgctg ccagtccccactgaggaagg aaaccccctc ctcaaacact 2160 atcgggggcc cgcaggggat gccacggtcgcctctgagaa ggaatcagtc atgtaaaccc 2220 cgggagggac cttccctgcc ctgctgggggtgctctttgg acactggatt atgaggaatg 2280 gataaatgga tgagctaggg ctctgggggtctgcctgcac actctgggga gccaggggcc 2340 ccagcaccct ccaggacagg agatctgggatgcctggctg ctggagtaca tgtgttcaca 2400 agggttactc ctcaaaaccc ccagttctcactcatgtccc caactcaagg ctagaaaaca 2460 gcaagatgga gaaataatgt tctgctgcgtccccaccgtg acctgcctgg cctcccctgt 2520 ctcagggagc aggtcacagg tcaccatggggaattctagc ccccactggg gggatgttac 2580 aacaccatgc tggttatttt ggcggctgtagttgtggggg gatgtgtgtg tgcacgtgtg 2640 tgtgtgtgtg tgtgtgtgtg tgtgtgtgtgttctgtgacc tcctgtcccc atggtacgtc 2700 ccaccctgtc cccagatccc ctattccctccacaataaca gaaacactcc cagggactct 2760 ggggagaggc tgaggacaaa tacctgctgtcactccagag gacatttttt ttagcaataa 2820 aattgagtgt caactattta aaaaaaaaaaaaaaaa 2856 8 2646 DNA Homo sapiens 8 tttccttcct cttttggcaa catggcgggcggagaagctg gagtgactct agggcagccg 60 catctttcgc gtcaggatct caccaccttggatgttacca agttgacgcc actttcacac 120 gaagttatca gcagacaagc cacaattaacataggtacaa ttggtcatgt agctcatggg 180 aaatccacag tcgtcaaagc tatttctggagttcatactg tcaggttcaa aaatgaacta 240 gaaagaaata ttacaatcaa gcttggatatgctaatgcta agatttataa gcttgatgac 300 ccaagttgcc ctcggccaga atgttatagatcttgtggga gcagtacacc tgacgagttt 360 cctacggaca ttccagggac caaagggaacttcaaattag tcagacatgt ttcctttgtt 420 gactgtcctg gccacgatat tttgatggctactatgctga acggtgcagc agtgatggat 480 gcagctcttc tgttgatagc tggtaatgaatcttgccctc agcctcagac atcggaacac 540 ctggctgcta tagagatcat gaaactgaagcatattttga ttctacaaaa taaaattgat 600 ttggtaaaag aaagtcaggc taaagaacaatacgagcaga tccttgcatt tgtccaaggt 660 acagtagcag agggagctcc cattattccaatttcagctc agctgaaata caatattgaa 720 gttgtttgtg agtacatagt aaagaaaattccagtacccc caagagactt tacttcagag 780 ccccggctta ttgttattag atcttttgatgtcaacaaac ctggctgtga agttgatgac 840 cttaagggag gtgtagctgg tggtagtatcctaaaaggag tattaaaggt gggccaggag 900 atagaagtaa gacctggtat tgtttccaaagatagtgaag gaaaactcat gtgtaaacca 960 atcttttcca aaattgtatc actttttgcggagcataatg atctgcaata tgctgctcca 1020 ggcggtctta ttggagttgg aacaaaaattgaccccactt tgtgccgggc tgacagaatg 1080 gtggggcaag tacttggtgc agtcggagctttacctgaga tattcacaga attggaaatt 1140 tcctatttcc tgcttagacg gcttctaggtgtacgcactg aaggagacaa gaaagcagca 1200 aaggttcaaa agctgtctaa gaatgaagtgctcatggtga acataggatc cctgtcaaca 1260 ggagggagag ttagtgctgt caaggccgatttgggtaaaa ttgttttgac caatccagtg 1320 tgcacagagg taggagaaaa aattgcccttagccgaagag ttgaaaaaca ctggcgttta 1380 attggttggg gtcagataag aagaggagtgacaatcaagc caacagtaga tgatgactga 1440 agaataccag ttaaataata cattcggatggatttggaag ttggaattcc tcttaacaac 1500 caaggggttt attttcaaag caatattggggaattgattt cacagttcgt taccttagta 1560 ggtaacggta aggttattct ctttttttttttttttggtt atgaaaactt agggactaaa 1620 attaatataa aaattggcat aatgttggattgaatctaca ttttggcaga agttaaacat 1680 tcccacataa tgtcaaaatt atacatcatgcagttctgtt tttttgtttg tttaattttg 1740 ttttgttttt gagtctggct ctgtcacccaggctggagtg cagtggcgtg atctgcaacc 1800 tctgcccccc gggttcaagc gattctcctgcctcagcctc ccgagtagct gagattacag 1860 gtgcgcgcca ccacacttgg ctaatttttgtattattagt agagacgggg tttcagcatg 1920 ttggctaggc cggtctctcc tgacctcagggtgatcagcc cacctcggcc tcacaaagtg 1980 ctgggattac aggcgtgagc caccttgcccagcccacatc atacagtttg aaatgaaact 2040 ttgccacaac cagcctttgc tgtagcacacacatatatca ctgaacctgt ttgaaataaa 2100 gttttttttc tttttcatga ttcgtctttgagtacctcca ggctgaaaga ctgttgtacc 2160 agtaaaaact taaaggcaca aattctccttgaagaccttc tcccttttat gtggccccat 2220 attttatgtt gctttatctt tgaaattttgcatgaaaagg aaatgaatgg attcgaatga 2280 aattgtcctt tagagcatga ttacttgttcccatggacaa atatttttct ccccttgctc 2340 ttcctggcct gaaacacggg aaaccagagtcaaaagttat ctccctctcc ctgtgatgcc 2400 ttgagatttt tttctgcgtt gtttaatgcctgaaatccaa gtcttcctcc atgggaaaat 2460 actgttatac caaataattc tagatgagtaacaaagatct ttttaggcct tcattttatg 2520 ttttttctta actgttatat tatgattgtgacatagatta tactactact aatttttgga 2580 tgtttcaaaa ggtcaagaag taaaagatgttagaaagcaa aaaaaaaaaa aaaaaaaaaa 2640 aaaaaa 2646 9 3151 DNA Homosapiens 9 ccggccagcg ggcgggctcc ccagccaggc cgctgcacct gtcaggggaacaagctggag 60 gagcaggacc ctagacctct gcagcccata ccaggtctca tggaggggaacaagctggag 120 gagcaggact ctagccctcc acagtccact ccagggctca tgaaggggaacaagcgtgag 180 gagcaggggc tgggccccga acctgcggcg ccccagcagc ccacggcggaggaggaggcc 240 ctgatcgagt tccaccgctc ctaccgagag ctcttcgagt tcttctgcaacaacaccacc 300 atccacggcg ccatccgcct ggtgtgctcc cagcacaacc gcatgaagacggccttctgg 360 gcagtgctgt ggctctgcac ctttggcatg atgtactggc aattcggcctgcttttcgga 420 gagtacttca gctaccccgt cagcctcaac atcaacctca actcggacaagctcgtcttc 480 cccgcagtga ccatctgcac cctcaatccc tacaggtacc cggaaattaaagaggagctg 540 gaggagctgg accgcatcac agagcagacg ctctttgacc tgtacaaatacagctccttc 600 accactctcg tggccggctc ccgcagccgt cgcgacctgc gggggactctgccgcacccc 660 ttgcagcgcc tgagggtccc gcccccgcct cacggggccc gtcgagcccgtagcgtggcc 720 tccagcttgc gggacaacaa cccccaggtg gactggaagg actggaagatcggcttccag 780 ctgtgcaacc agaacaaatc ggactgcttc taccagacat actcatcaggggtggatgcg 840 gtgagggagt ggtaccgctt ccactacatc aacatcctgt cgaggctgccagagactctg 900 ccatccctgg aggaggacac gctgggcaac ttcatcttcg cctgccgcttcaaccaggtc 960 tcctgcaacc aggcgaatta ctctcacttc caccacccga tgtatggaaactgctatact 1020 ttcaatgaca agaacaactc caacctctgg atgtcttcca tgcctggaatcaacaacggt 1080 ctgtccctga tgctgcgcgc agagcagaat gacttcattc ccctgctgtccacagtgact 1140 ggggcccggg taatggtgca cgggcaggat gaacctgcct ttatggatgatggtggcttt 1200 aacttgcggc ctggcgtgga gacctccatc agcatgagga aggaaaccctggacagactt 1260 gggggcgatt atggcgactg caccaagaat ggcagtgatg ttcctgttgagaacctttac 1320 ccttcaaagt acacacagca ggtgtgtatt cactcctgct tccaggagagcatgatcaag 1380 gagtgtggct gtgcctacat cttctatccg cggccccaga acgtggagtactgtgactac 1440 agaaagcaca gttcctgggg gtactgctac tataagctcc aggttgacttctcctcagac 1500 cacctgggct gtttcaccaa gtgccggaag ccatgcagcg tgaccagctaccagctctct 1560 gctggttact cacgatggcc ctcggtgaca tcccaggaat gggtcttccagatgctatcg 1620 cgacagaaca attacaccgt caacaacaag agaaatggag tggccaaagtcaacatcttc 1680 ttcaaggagc tgaactacaa aaccaattct gagtctccct ctgtcacgatggtcaccctc 1740 ctgtccaacc tgggcagcca gtggagcctg tggttcggct cctcggtgttgtctgtggtg 1800 gagatggctg agctcgtctt tgacctgctg gtcatcatgt tcctcatgctgctccgaagg 1860 ttccgaagcc gatactggtc tccaggccga gggggcaggg gtgctcaggaggtagcctcc 1920 accctggcat cctcccctcc ttcccacttc tgcccccacc ccatgtctctgtccttgtcc 1980 cagccaggcc ctgctccctc tccagccttg acagcccctc cccctgcctatgccaccctg 2040 ggcccccgcc catctccagg gggctctgca ggggccagtt cctccacctgtcctctgggg 2100 gggccctgag agggaaggag aggtttctca caccaaggca gatgctcctctggtgggagg 2160 gtgctggccc tggcaagatt gaaggatgtg cagggcttcc tctcagagccgcccaaactg 2220 ccgttgatgt gtggagggga agcaagatgg gtaagggctc aggaagttgctccaagaaca 2280 gtagctgatg aagctgccca gaagtgcctt ggctccagcc ctgtaccccttggtactgcc 2340 tctgaacact ctggtttccc cacccaactg cggctaagtc tctttttcccttggatcagc 2400 caagcgaaac ttggagcttt gacaaggaac tttcctaaga aaccgctgataaccaggaca 2460 aaacacaacc aagggtacac gcaggcatgc acgggtttcc tgcccagcgacggcttaagc 2520 cagcccccga ctggcctggc cacactgctc tccagtagca cagatgtctgctcctcctct 2580 tgaacttggg tgggaaaccc cacccaaaag ccccctttgt tacttaggcaattccccttc 2640 cctgactccc gagggctagg gctagagcag acccgggtaa gtaaaggcagacccagggct 2700 cctctagcct catacccgtg ccctcacaga gccatgcccc ggcacctctgccctgtgtct 2760 ttcatacctc tacatgtctg cttgagatat ttcctcagcc tgaaagtttccccaaccatc 2820 tgccagagaa ctcctatgca tcccttagaa ccctgctcag acaccattacttttgtgaac 2880 gcttctgcca catcttgtct tccccaaaat tgatcactcc gccttctcctgggctcccgt 2940 agcacactat aacatctgct ggagtgttgc tgttgcacca tactttcttgtacatttgtg 3000 tctcccttcc caactagact gtaagtgcct tgcggtcagg gactgaatcttgcccgttta 3060 tgtatgctcc atgtctagcc catcatcctg cttggagcaa gtaggcaggagctcaataaa 3120 tgtttgttgc atgaaaaaaa aaaaaaaaaa a 3151 10 4050 DNA Homosapiens 10 cttgcaatcc aggctttcct tggaagtggc tgtaacatgt atgaaaagaaagaaaggagg 60 accaagagat gaaagagggc tgcacgcgtg ggggcccgag tggtgggcggggacagtcgt 120 cttgttacag gggtgctggc cttccctggc gcctgcccct gtcggccccgcccgagaacc 180 tccctgcgcc agggcagggt ttactcatcc cggcgaggtg atcccatgcgcgagggcggg 240 cgcaagggcg gccagagaac ccagcaatcc gagtatgcgg catcagcccttcccaccagg 300 cacttccttc cttttcccga acgtccaggg agggagggcc gggcacttataaactcgagc 360 cctggccgat ccgcatgtca gaggctgcct cgcaggggct gcgcgcacggcaagaagtgt 420 ctgggctggg acggacagga gaggctgtcg ccatcggcgt cctgtgcccctctgctccgg 480 cacggccctg tcgcagtgcc cgcgctttcc ccggcgcctg cacgcggcgcgcctgggtaa 540 catgcttggg gtcctggtcc ttggcgcgct ggccctggcc ggcctggggttccccgcacc 600 cgcagagccg cagccgggtg gcagccagtg cgtcgagcac gactgcttcgcgctctaccc 660 gggccccgcg accttcctca atgccagtca gatctgcgac ggactgcggggccacctaat 720 gacagtgcgc tcctcggtgg ctgccgatgt catttccttg ctactgaacggcgacggcgg 780 cgttggccgc cggcgcctct ggatcggcct gcagctgcca cccggctgcggcgaccccaa 840 gcgcctcggg cccctgcgcg gcttccagtg ggttacggga gacaacaacaccagctatag 900 caggtgggca cggctcgacc tcaatggggc tcccctctgc ggcccgttgtgcgtcgctgt 960 ctccgctgct gaggccactg tgcccagcga gccgatctgg gaggagcagcagtgcgaagt 1020 gaaggccgat ggcttcctct gcgagttcca cttcccagcc acctgcaggccactggctgt 1080 ggagcccggc gccgcggctg ccgccgtctc gatcacctac ggcaccccgttcgcggcccg 1140 cggagcggac ttccaggcgc tgccggtggg cagctccgcc gcggtggctcccctcggctt 1200 acagctaatg tgcaccgcgc cgcccggagc ggtccagggg cactgggccagggaggcgcc 1260 gggcgcttgg gactgcagcg tggagaacgg cggctgcgag cacgcgtgcaatgcgatccc 1320 tggggctccc cgctgccagt gcccagccgg cgccgccctg caggcagacgggcgctcctg 1380 caccgcatcc gcgacgcagt cctgcaacga cctctgcgag cacttctgcgttcccaaccc 1440 cgaccagccg ggctcctact cgtgcatgtg cgagaccggc taccggctggcggccgacca 1500 acaccggtgc gaggacgtgg atgactgcat actggagccc agtccgtgtccgcagcgctg 1560 tgtcaacaca cagggtggct tcgagtgcca ctgctaccct aactacgacctggtggacgg 1620 cgagtgtgtg gagcccgtgg acccgtgctt cagagccaac tgcgagtaccagtgccagcc 1680 cctgaaccaa actagctacc tctgcgtctg cgccgagggc ttcgcgcccattccccacga 1740 gccgcacagg tgccagatgt tttgcaacca gactgcctgt ccagccgactgcgaccccaa 1800 cacccaggct agctgtgagt gccctgaagg ctacatcctg gacgacggtttcatctgcac 1860 ggacatcgac gagtgcgaaa acggcggctt ctgctccggg gtgtgccacaacctccccgg 1920 taccttcgag tgcatctgcg ggcccgactc ggcccttgcc cgccacattggcaccgactg 1980 tgactccggc aaggtggacg gtggcgacag cggctctggc gagcccccgcccagcccgac 2040 gcccggctcc accttgactc ctccggccgt ggggctcgtg cattcgggcttgctcatagg 2100 catctccatc gcgagcctgt gcctggtggt ggcgcttttg gcgctcctctgccacctgcg 2160 caagaagcag ggcgccgcca gggccaagat ggagtacaag tgcgcggccccttccaagga 2220 ggtagtgctg cagcacgtgc ggaccgagcg gacgccgcag agactctgagcggcctccgt 2280 ccaggagcct ggctccgtcc aggagctgtg cctcctcacc cccagctttgctaccaaagc 2340 accttagctg gcattacagc tggagaagac cctccccgca ccccccaagctgttttcttc 2400 tattccatgg ctaactggcg agggggtgat tagagggagg agaatgagcctcggcctctt 2460 ccgtgacgtc actggaccac tgggcaatga tggcaatttt gtaacgaagacacagactgc 2520 gatttgtccc aggtcctcac taccgggcgc aggagggtga gcgttattggtcggcagcct 2580 tctgggcaga ccttgacctc gtgggctagg gatgactaaa atatttattttttttaagta 2640 tttaggtttt tgtttgtttc ctttgttctt acctgtatgt ctccagtatccactttgcac 2700 agctctccgg tctctctctc tctacaaact cccacttgtc atgtgacaggtaaactatct 2760 tggtgaattt ttttttccta gccctctcac atttatgaag caagccccacttattcccca 2820 ttcttcctag ttttctcctc ccaggaactg ggccaactca cctgagtcaccctacctgtg 2880 cctgacccta cttcttttgc tcatctagct gtctgctcag acagaacccctacatgaaac 2940 agaaacaaaa acactaaaaa taaaaatggc catttgcttt ttcaccagatttgctaattt 3000 atcctgaaat ttcagattcc cagagcaaaa taattttaaa caaagggttgagatgtaaaa 3060 ggtattaaat tgatgttgct ggactgtcat agaaattaca cccaaagaggtatttatctt 3120 tacttttaaa cagtgagcct gaattttgtt gctgttttga tttgtactgaaaaatggtaa 3180 ttgttgctaa tcttcttatg caatttcctt ttttgttatt attacttatttttgacagtg 3240 ttgaaaatgt tcagaaggtt gctctagatt gagagaagag acaaacacctcccaggagac 3300 agttcaagaa agcttcaaac tgcatgattc atgccaatta gcaattgactgtcactgttc 3360 cttgtcactg gtagaccaaa ataaaaccag ctctactggt cttgtggaattgggagcttg 3420 ggaatggatc ctggaggatg cccaattagg gcctagcctt aatcaggtcctcagagaatt 3480 tctaccattt cagagaggcc ttttggaatg tggcccctga acaagaattggaagctgccc 3540 tgcccatggg agctggttag aaatgcagaa tcctaggctc caccccatccagttcatgag 3600 aatctatatt taacaagatc tgcagggggt gtgtctgctc agtaatttgaggacaaccat 3660 tccagactgc ttccaatttt ctggaataca tgaaatatag atcagttataagtagcaggc 3720 caagtcaggc ccttattttc aagaaactga ggaattttct ttgtgtagctttgctctttg 3780 gtagaaaagg ctaggtacac agctctagac actgccacac agggtctgcaaggtctttgg 3840 ttcagctaag ctaggaatga aatcctgctt cagtgtatgg aaataaatgtatcatagaaa 3900 tgtaactttt gtaagacaaa ggttttcctc ttctattttg taaactcaaaatatttgtac 3960 atagttattt atttattgga gataatctag aacacaggca aaatccttgcttatgacatc 4020 acttgtacaa aataaacaaa taacaatgtg 4050 11 4465 DNA Homosapiens 11 caattgtcat acgacttgca gtgagcgtca ggagcacgtc caggaactcctcagcagcgc 60 ctccttcagc tccacagcca gacgccctca gacagcaaag cctacccccgcgccgcgccc 120 tgcccgccgc tcggatgctc gcccgcgccc tgctgctgtg cgcggtcctggcgctcagcc 180 atacagcaaa tccttgctgt tcccacccat gtcaaaaccg aggtgtatgtatgagtgtgg 240 gatttgacca gtataagtgc gattgtaccc ggacaggatt ctatggagaaaactgctcaa 300 caccggaatt tttgacaaga ataaaattat ttctgaaacc cactccaaacacagtgcact 360 acatacttac ccacttcaag ggattttgga acgttgtgaa taacattcccttccttcgaa 420 atgcaattat gagttatgtc ttgacatcca gatcacattt gattgacagtccaccaactt 480 acaatgctga ctatggctac aaaagctggg aagccttctc taacctctcctattatacta 540 gagcccttcc tcctgtgcct gatgattgcc cgactccctt gggtgtcaaaggtaaaaagc 600 agcttcctga ttcaaatgag attgtggaaa aattgcttct aagaagaaagttcatccctg 660 atccccaggg ctcaaacatg atgtttgcat tctttgccca gcacttcacgcatcagtttt 720 tcaagacaga tcataagcga gggccagctt tcaccaacgg gctgggccatggggtggact 780 taaatcatat ttacggtgaa actctggcta gacagcgtaa actgcgccttttcaaggatg 840 gaaaaatgaa atatcagata attgatggag agatgtatcc tcccacagtcaaagatactc 900 aggcagagat gatctaccct cctcaagtcc ctgagcatct acggtttgctgtggggcagg 960 aggtctttgg tctggtgcct ggtctgatga tgtatgccac aatctggctgcgggaacaca 1020 acagagtatg cgatgtgctt aaacaggagc atcctgaatg gggtgatgagcagttgttcc 1080 agacaagcag gctaatactg ataggagaga ctattaagat tgtgattgaagattatgtgc 1140 aacacttgag tggctatcac ttcaaactga aatttgaccc agaactacttttcaacaaac 1200 aattccagta ccaaaatcgt attgctgctg aatttaacac cctctatcactggcatcccc 1260 ttctgcctga cacctttcaa attcatgacc agaaatacaa ctatcaacagtttatctaca 1320 acaactctat attgctggaa catggaatta cccagtttgt tgaatcattcaccaggcaaa 1380 ttgctggcag ggttgctggt ggtaggaatg ttccacccgc agtacagaaagtatcacagg 1440 cttccattga ccagagcagg cagatgaaat accagtcttt taatgagtaccgcaaacgct 1500 ttatgctgaa gccctatgaa tcatttgaag aacttacagg agaaaaggaaatgtctgcag 1560 agttggaagc actctatggt gacatcgatg ctgtggagct gtatcctgcccttctggtag 1620 aaaagcctcg gccagatgcc atctttggtg aaaccatggt agaagttggagcaccattct 1680 ccttgaaagg acttatgggt aatgttatat gttctcctgc ctactggaagccaagcactt 1740 ttggtggaga agtgggtttt caaatcatca acactgcctc aattcagtctctcatctgca 1800 ataacgtgaa gggctgtccc tttacttcat tcagtgttcc agatccagagctcattaaaa 1860 cagtcaccat caatgcaagt tcttcccgct ccggactaga tgatatcaatcccacagtac 1920 tactaaaaga acgttcgact gaactgtaga agtctaatga tcatatttatttatttatat 1980 gaaccatgtc tattaattta attatttaat aatatttata ttaaactccttatgttactt 2040 aacatcttct gtaacagaag tcagtactcc tgttgcggag aaaggagtcatacttgtgaa 2100 gacttttatg tcactactct aaagattttg ctgttgctgt taagtttggaaaacagtttt 2160 tattctgttt tataaaccag agagaaatga gttttgacgt ctttttacttgaatttcaac 2220 ttatattata agaacgaaag taaagatgtt tgaatactta aacactatcacaagatggca 2280 aaatgctgaa agtttttaca ctgtcgatgt ttccaatgca tcttccatgatgcattagaa 2340 gtaactaatg tttgaaattt taaagtactt ttggttattt ttctgtcatcaaacaaaaac 2400 aggtatcagt gcattattaa atgaatattt aaattagaca ttaccagtaatttcatgtct 2460 actttttaaa atcagcaatg aaacaataat ttgaaatttc taaattcatagggtagaatc 2520 acctgtaaaa gcttgtttga tttcttaaag ttattaaact tgtacatataccaaaaagaa 2580 gctgtcttgg atttaaatct gtaaaatcag atgaaatttt actacaattgcttgttaaaa 2640 tattttataa gtgatgttcc tttttcacca agagtataaa cctttttagtgtgactgtta 2700 aaacttcctt ttaaatcaaa atgccaaatt tattaaggtg gtggagccactgcagtgtta 2760 tctcaaaata agaatatttt gttgagatat tccagaattt gtttatatggctggtaacat 2820 gtaaaatcta tatcagcaaa agggtctacc tttaaaataa gcaataacaaagaagaaaac 2880 caaattattg ttcaaattta ggtttaaact tttgaagcaa acttttttttatccttgtgc 2940 actgcaggcc tggtactcag attttgctat gaggttaatg aagtaccaagctgtgcttga 3000 ataacgatat gttttctcag attttctgtt gtacagttta atttagcagtccatatcaca 3060 ttgcaaaagt agcaatgacc tcataaaata cctcttcaaa atgcttaaattcatttcaca 3120 cattaatttt atctcagtct tgaagccaat tcagtaggtg cattggaatcaagcctggct 3180 acctgcatgc tgttcctttt cttttcttct tttagccatt ttgctaagagacacagtctt 3240 ctcatcactt cgtttctcct attttgtttt actagtttta agatcagagttcactttctt 3300 tggactctgc ctatattttc ttacctgaac ttttgcaagt tttcaggtaaacctcagctc 3360 aggactgcta tttagctcct cttaagaaga ttaaaagaga aaaaaaaaggcccttttaaa 3420 aatagtatac acttatttta agtgaaaagc agagaatttt atttatagctaattttagct 3480 atctgtaacc aagatggatg caaagaggct agtgcctcag agagaactgtacggggtttg 3540 tgactggaaa aagttacgtt cccattctaa ttaatgccct ttcttatttaaaaacaaaac 3600 caaatgatat ctaagtagtt ctcagcaata ataataatga cgataatacttcttttccac 3660 atctcattgt cactgacatt taatggtact gtatattact taatttattgaagattatta 3720 tttatgtctt attaggacac tatggttata aactgtgttt aagcctacaatcattgattt 3780 ttttttgtta tgtcacaatc agtatatttt ctttggggtt acctctctgaatattatgta 3840 aacaatccaa agaaatgatt gtattaagat ttgtgaataa atttttagaaatctgattgg 3900 catattgaga tatttaaggt tgaatgtttg tccttaggat aggcctatgtgctagcccac 3960 aaagaatatt gtctcattag cctgaatgtg ccataagact gaccttttaaaatgttttga 4020 gggatctgtg gatgcttcgt taatttgttc agccacaatt tattgagaaaatattctgtg 4080 tcaagcactg tgggttttaa tatttttaaa tcaaacgctg attacagataatagtattta 4140 tataaataat tgaaaaaaat tttcttttgg gaagagggag aaaatgaaataaatatcatt 4200 aaagataact caggagaatc ttctttacaa ttttacgttt agaatgtttaaggttaagaa 4260 agaaatagtc aatatgcttg tataaaacac tgttcactgt tttttttaaaaaaaaaactt 4320 gatttgttat taacattgat ctgctgacaa aacctgggaa tttgggttgtgtatgcgaat 4380 gtttcagtgc ctcagacaaa tgtgtattta acttatgtaa aagataagtctggaaataaa 4440 tgtctgttta tttttgtact attta 4465 12 1856 DNA Homosapiens 12 gggagaaccg ttcgcggagg aaaggcgaac tagtgttggg atggccaccaactgggggag 60 cctcttgcag gataaacagc agctagagga gctggcacgg caggccgtggaccgggccct 120 ggctgaggga gtattgctga ggacctcaca ggagcccact tcctcggaggtggtgagcta 180 tgccccattc acgctcttcc cctcactggt ccccagtgcc ctgctggagcaagcctatgc 240 tgtgcagatg gacttcaacc tgctagtgga tgctgtcagc cagaacgctgccttcctgga 300 gcaaactctt tccagcacca tcaaacagga tgactttacc gctcgtctctttgacatcca 360 caagcaagtc ctaaaagagg gcattgccca gactgtgttc ctgggcctgaatcgctcaga 420 ctacatgttc cagcgcagcg cagatggctc cccagccctg aaacagatcgaaatcaacac 480 catctctgcc agctttgggg gcctggcctc ccggacccca gctgtgcaccgacatgttct 540 cagtgtcctg agtaagacca aagaagctgg caagatcctc tctaataatcccagcaaggg 600 actggccctg ggaattgcca aagcctggga gctctacggc tcacccaatgctctggtgct 660 actgattgct caagagaagg aaagaaacat atttgaccag cgtgccatagagaatgagct 720 actggccagg aacatccatg tgatccgacg aacatttgaa gatatctctgaaaaggggtc 780 tctggaccaa gaccgaaggc tgtttgtgga tggccaggaa attgctgtggtttacttccg 840 ggatggctac atgcctcgtc agtacagtct acagaattgg gaagcacgtctactgctgga 900 gaggtcacat gctgccaagt gcccagacat tgccacccag ctggctgggactaagaaggt 960 gcagcaggag ctaagcaggc cgggcatgct ggagatgttg ctccctggccagcctgaggc 1020 tgtggcccgc ctccgcgcca cctttgctgg cctctactca ctggatgtgggtgaagaagg 1080 ggaccaggcc atcgccgagg cccttgctgc ccctagccgg tttgtgctaaagccccagag 1140 agagggtgga ggtaacaacc tatatgggga ggaaatggta caggccctgaaacagctgaa 1200 ggacagtgag gagagggcct cctacatcct catggagaag atcgaacctgagccttttga 1260 gaattgcctg ctacggcctg gcagccctgc ccgagtggtc cagtgcatttcagagctggg 1320 catctttggg gtctatgtca ggcaggaaaa gacactcgtg atgaacaagcacgtggggca 1380 tctacttcga accaaagcca tcgagcatgc agatggtggt gtggcagcgggagtggcagt 1440 cctggacaac ccataccctg tgtgagggca caaccaggcc acgggaccttctatcctctg 1500 tatttgtcat tcctctccta gccctcctga ggggtatcct cctaaagacctccaaagttt 1560 ttatggaagg gtaaatactg gtaccttccc ccagctttcc atctgaggaccagaaaagtt 1620 gtgtctccct tagatgagat ctagacgccc ccaaatcctt gagatgtgggtatagctcag 1680 ggtaagctgc tctgaggtaa aggtccatga accctgcccc actcctgtcagcccctcatc 1740 agccttttca gcaggttcca gtgcctgact tgggatagga ctgagtggtaggaggagggg 1800 gagtggaggg gcatagcctt tccctaattc tgccttaaat aaaactgcattgctgt 1856 13 2473 DNA Homo sapiens 13 aatcgcgaaa cccggcgagc ggcgcgctggctatcgagcg agcggggcgg aaccgggagt 60 tgcgccgccg ctcgggcgcc gggctccgtcgcggccgcag ccccgcgggt cgccctcccg 120 tgcctcgccc gcggacaccc tggccgtggacaccctggcc gtgggcaccc gcggggcgcg 180 gcgcgggcgc tgcgcggcgg cggcggcggcatgaaggtca cgtcgctcga cgggcgccag 240 ctgcgcaaga tgctccgcaa ggaggcggcggcgcgctgcg tggtgctcga ctgccggccc 300 tatctggcct tcgctgcctc gaacgtgcgcggctcgctca acgtcaacct caactcggtg 360 gtgctgcggc gggcccgggg cggcgcggtgtcggcgcgct acgtgctgcc cgacgaggcg 420 gcgcgcgcgc ggctcctgca ggagggcggcggcggcgtcg cggccgtggt ggtgctggac 480 cagggcagcc gccactggca gaagctgcgagaggagagcg ccgcgcgtgt cgtcctcacc 540 tcgctactcg cttgcctacc cgccggcccgcgggtctact tcctcaaagg gggatatgag 600 actttctact cggaatatcc tgagtgttgcgtggatgtaa aacccatttc acaagagaag 660 attgagagtg agagagccct catcagccagtgtggaaaac cagtggtaaa tgtcagctac 720 aggccagctt atgaccaggg tggcccagttgaaatccttc ccttcctcta ccttggaagt 780 gcctaccatg catccaagtg cgagttcctcgccaacttgc acatcacagc cctgctgaat 840 gtctcccgac ggacctccga ggcctgcatgacccacctac actacaaatg gatccctgtg 900 gaagacagcc acacggctga cattagctcccactttcaag aagcaataga cttcattgac 960 tgtgtcaggg aaaagggagg caaggtcctggtccactgtg aggctgggat ctcccgttca 1020 cccaccatct gcatggctta ccttatgaagaccaagcagt tccgcctgaa ggaggccttc 1080 gattacatca agcagaggag gagcatggtctcgcccaact ttggcttcat gggccagctc 1140 ctgcagtacg aatctgagat cctgccctccacgcccaacc cccagcctcc ctcctgccaa 1200 ggggaggcag caggctcttc actgataggccatttgcaga cactgagccc tgacatgcag 1260 ggtgcctact gcacattccc tgcctcggtgctggcaccgg tgcctaccca ctcaacagtc 1320 tcagagctca gcagaagccc tgtggcaacggccacatcct gctaaaactg ggatggagga 1380 atcggcccag ccccaagagc aactgtgatttttgttttta agactcatgg acatttcata 1440 cctgtgcaat actgaagacc tcattctgtcatgctgcccc agtgagatag tgagtggtca 1500 ccaggcttgc aaatgaactt cagacggacctcagggtagg ttctcgggac tgaaggaagg 1560 ccaagccatt acgggagcac agcatgtgctgactactgta cttccagacc cctgccctct 1620 tgggactgcc cagtccttgc acctcagagttcgccttttc atttcaagca taagccaata 1680 aatacctgca gcaacgtggg agaaagaagttgctggacca ggagaaaagg cagttatgaa 1740 gccaattcat tttgaaggaa gcacaatttccaccttattt tttgaacttt ggcagtttca 1800 atgtctgtct ctgttgcttc ggggcataagctgatcaccg tctagttggg aaagtcaccc 1860 tacagggttt gtagggacat gatcagcatcctgatttgaa ccctgaaatg ttgtgtagac 1920 accctcttgg gtccaatgag gtagttggttgaagtagcaa gatgttggct tttctggatt 1980 ttttttgcca tgggttcttc actgaccttggactttggca tgattcttag tcatacttga 2040 acttgtctca ttccacctct tctcagagcaactcttcctt tgggaaaaga gttcttcaga 2100 tcatagacca aaaaagtcat accttcgaggtggtagcagt agattccagg aggagaaggg 2160 tacttgctag gtatcctggg tcagtggcggtgcaaactgg tttcctcagc tgcctgtcct 2220 tctgtgtgct tatgtctctt gtgacaattgttttcctccc tgcccctgga ggttgtcttc 2280 aactgtggac ttctgggatt tgcagattttgcaacgtggt actacttttt tttctttttg 2340 tctgttagtt atttctccag gggaaaaggcaataattttc taagacccgt gtgaatgtga 2400 agaaaagcag tatgttactg gttgttgttgttgttcttgt tttttatatg taaaataaaa 2460 atagtgaaag gag 2473 14 976 DNAHomo sapiens 14 cccggaacct ggcgcaactc ctagagcggt ccttggggag acgcgggtcccagtcctgcg 60 gctcctactg gggagtgcgc tggtcggaag attgctggac tcgctgaagagagactacgc 120 aggaaagccc cagccaccca tcaaatcaga gagaaggaat ccaccttcttacgctatggc 180 aggtaagaaa gtactcattg tctatgcaca ccaggaaccc aagtctttcaacggatcctt 240 gaagaatgtg gctgtagatg aactgagcag gcagggctgc accgtcacagtgtctgattt 300 gtatgccatg aactttgagc cgagggccac agacaaagat atcactggtactctttctaa 360 tcctgaggtt ttcaattatg gagtggaaac ccacgaagcc tacaagcaaaggtctctggc 420 tagcgacatc actgatgagc agaaaaaggt tcgggaggct gacctagtgatatttcagtt 480 cccgctgtac tggttcagcg tgccggccat cctgaagggc tggatggatagggtgctgtg 540 ccagggcttt gcctttgaca tcccaggatt ctacgattcc ggtttgctccagggtaaact 600 agcgctcctt tccgtaacca cgggaggcac ggccgagatg tacacgaagacaggagtcaa 660 tggagattct cgatacttcc tgtggccact ccagcatggc acattacacttctgtggatt 720 taaagtcctt gcccctcaga tcagctttgc tcctgaaatt gcatccgaagaagaaagaaa 780 ggggatggtg gctgcgtggt cccagaggct gcagaccatc tggaaggaagagcccatccc 840 ctgcacagcc cactggcact tcgggcaata actctgtggc acgtgggcatcacgtaagca 900 gcacactagg aggcccaggc gcaggcaaag agaagatggt gctgtcatgaaataaaatta 960 caacatagct acctgg 976 15 7560 DNA Homo sapiens 15accggccaca gcctgcctac tgtcacccgc ctctcccgcg cgcagataca cgcccccgcc 60tccgtgggca caaaggcagc gctgctgggg aactcggggg aacgcgcacg tgggaaccgc 120cgcagctcca cactccaggt acttcttcca aggacctagg tctctcgccc atcggaaaga 180aaataattct ttcaagaaga tcagggacaa ctgatttgaa gtctactctg tgcttctaaa 240tccccaattc tgctgaaagt gaatccctag agccctagag ccccagcagc acccagccaa 300acccacctcc accatggggg ccatgactca gctgttggca ggtgtctttc ttgctttcct 360tgccctcgct accgaaggtg gggtcctcaa gaaagtcatc cggcacaagc gacagagtgg 420ggtgaacgcc accctgccag aagagaacca gccagtggtg tttaaccacg tttacaacat 480caagctgcca gtgggatccc agtgttcggt ggatctggag tcagccagtg gggagaaaga 540cctggcaccg ccttcagagc ccagcgaaag ctttcaggag cacacagtag atggggaaaa 600ccagattgtc ttcacacatc gcatcaacat cccccgccgg gcctgtggct gtgccgcagc 660ccctgatgtt aaggagctgc tgagcagact ggaggagctg gagaacctgg tgtcttccct 720gagggagcaa tgtactgcag gagcaggctg ctgtctccag cctgccacag gccgcttgga 780caccaggccc ttctgtagcg gtcggggcaa cttcagcact gaaggatgtg gctgtgtctg 840cgaacctggc tggaaaggcc ccaactgctc tgagcccgaa tgtccaggca actgtcacct 900tcgaggccgg tgcattgatg ggcagtgcat ctgtgacgac ggcttcacgg gcgaggactg 960cagccagctg gcttgcccca gcgactgcaa tgaccagggc aagtgcgtga atggagtctg 1020catctgtttc gaaggctacg ccggggctga ctgcagccgt gaaatctgcc cagtgccctg 1080cagtgaggag cacggcacat gtgtagatgg cttgtgtgtg tgccacgatg gctttgcagg 1140cgatgactgc aacaagcctc tgtgtctcaa caattgctac aaccgtggac gatgcgtgga 1200gaatgagtgc gtgtgtgatg agggtttcac gggcgaagac tgcagtgagc tcatctgccc 1260caatgactgc ttcgaccggg gccgctgcat caatggcacc tgctactgcg aagaaggctt 1320cacaggtgaa gactgcggga aacccacctg cccacatgcc tgccacaccc agggccggtg 1380tgaggagggg cagtgtgtat gtgatgaggg ctttgccggt ttggactgca gcgagaagag 1440gtgtcctgct gactgtcaca atcgtggccg ctgtgtagac gggcggtgtg agtgtgatga 1500tggtttcact ggagctgact gtggggagct caagtgtccc aatggctgca gtggccatgg 1560ccgctgtgtc aatgggcagt gtgtgtgtga tgagggctat actggggagg actgcagcca 1620gctacggtgc cccaatgact gtcacagtcg gggccgctgt gtcgagggca aatgtgtatg 1680tgagcaaggc ttcaagggct atgactgcag tgacatgagc tgccctaatg actgtcacca 1740gcacggccgc tgtgtgaatg gcatgtgtgt ttgtgatgac ggctacacag gggaagactg 1800ccgggatcgc caatgcccca gggactgcag caacaggggc ctctgtgtgg acggacagtg 1860cgtctgtgag gacggcttca ccggccctga ctgtgcagaa ctctcctgtc caaatgactg 1920ccatggccag ggtcgctgtg tgaatgggca gtgcgtgtgc catgaaggat ttatgggcaa 1980agactgcaag gagcaaagat gtcccagtga ctgtcatggc cagggccgct gcgtggacgg 2040ccagtgcatc tgccacgagg gcttcacagg cctggactgt ggccagcact cctgccccag 2100tgactgcaac aacttaggac aatgcgtctc gggccgctgc atctgcaacg agggctacag 2160cggagaagac tgctcagagg tgtctcctcc caaagacctc gttgtgacag aagtgacgga 2220agagacggtc aacctggcct gggacaatga gatgcgggtc acagagtacc ttgtcgtgta 2280cacgcccacc cacgagggtg gtctggaaat gcagttccgt gtgcctgggg accagacgtc 2340caccatcatc caggagctgg agcctggtgt ggagtacttt atccgtgtat ttgccatcct 2400ggagaacaag aagagcattc ctgtcagcgc cagggtggcc acgtacttac ctgcacctga 2460aggcctgaaa ttcaagtcca tcaaggagac atctgtggaa gtggagtggg atcctctaga 2520cattgctttt gaaacctggg agatcatctt ccggaatatg aataaagaag atgagggaga 2580gatcaccaaa agcctgagga ggccagagac ctcttaccgg caaactggtc tagctcctgg 2640gcaagagtat gagatatctc tgcacatagt gaaaaacaat acccggggcc ctggcctgaa 2700gagggtgacc accacacgct tggatgcccc cagccagatc gaggtgaaag atgtcacaga 2760caccactgcc ttgatcacct ggttcaagcc cctggctgag atcgatggca ttgagctgac 2820ctacggcatc aaagacgtgc caggagaccg taccaccatc gatctcacag aggacgagaa 2880ccagtactcc atcgggaacc tgaagcctga cactgagtac gaggtgtccc tcatctcccg 2940cagaggtgac atgtcaagca acccagccaa agagaccttc acaacaggcc tcgatgctcc 3000caggaatctt cgacgtgttt cccagacaga taacagcatc accctggaat ggaggaatgg 3060caaggcagct attgacagtt acagaattaa gtatgccccc atctctggag gggaccacgc 3120tgaggttgat gttccaaaga gccaacaagc cacaaccaaa accacactca caggtctgag 3180gccgggaact gaatatggga ttggagtttc tgctgtgaag gaagacaagg agagcaatcc 3240agcgaccatc aacgcagcca cagagttgga cacgcccaag gaccttcagg tttctgaaac 3300tgcagagacc agcctgaccc tgctctggaa gacaccgttg gccaaatttg accgctaccg 3360cctcaattac agtctcccca caggccagtg ggtgggagtg cagcttccaa gaaacaccac 3420ttcctatgtc ctgagaggcc tggaaccagg acaggagtac aatgtcctcc tgacagccga 3480gaaaggcaga cacaagagca agcccgcacg tgtgaaggca tccactgaac aagcccctga 3540gctggaaaac ctcaccgtga ctgaggttgg ctgggatggc ctcagactca actggaccgc 3600ggctgaccag gcctatgagc actttatcat tcaggtgcag gaggccaaca aggtggaggc 3660agctcggaac ctcaccgtgc ctggcagcct tcgggctgtg gacataccgg gcctcaaggc 3720tgctacgcct tatacagtct ccatctatgg ggtgatccag ggctatagaa caccagtgct 3780ctctgctgag gcctccacag gggaaactcc caatttggga gaggtcgtgg tggccgaggt 3840gggctgggat gccctcaaac tcaactggac tgctccagaa ggggcctatg agtacttttt 3900cattcaggtg caggaggctg acacagtaga ggcagcccag aacctcaccg tcccaggagg 3960actgaggtcc acagacctgc ctgggctcaa agcagccact cattatacca tcaccatccg 4020cggggtcact caggacttca gcacaacccc tctctctgtt gaagtcttga cagaggaggt 4080tccagatatg ggaaacctca cagtgaccga ggttagctgg gatgctctca gactgaactg 4140gaccacgcca gatggaacct atgaccagtt tactattcag gtccaggagg ctgaccaggt 4200ggaagaggct cacaatctca cggttcctgg cagcctgcgt tccatggaaa tcccaggcct 4260cagggctggc actccttaca cagtcaccct gcacggcgag gtcaggggcc acagcactcg 4320accccttgct gtagaggtcg tcacagagga tctcccacag ctgggagatt tagccgtgtc 4380tgaggttggc tgggatggcc tcagactcaa ctggaccgca gctgacaatg cctatgagca 4440ctttgtcatt caggtgcagg aggtcaacaa agtggaggca gcccagaacc tcacgttgcc 4500tggcagcctc agggctgtgg acatcccggg cctcgaggct gccacgcctt atagagtctc 4560catctatggg gtgatccggg gctatagaac accagtactc tctgctgagg cctccacagc 4620caaagaacct gaaattggaa acttaaatgt ttctgacata actcccgaga gcttcaatct 4680ctcctggatg gctaccgatg ggatcttcga gacctttacc attgaaatta ttgattccaa 4740taggttgctg gagactgtgg aatataatat ctctggtgct gaacgaactg cccatatctc 4800agggctaccc cctagtactg attttattgt ctacctctct ggacttgctc ccagcatccg 4860gaccaaaacc atcagtgcca cagccacgac agaggccctg ccccttctgg aaaacctaac 4920catttccgac attaatccct acgggttcac agtttcctgg atggcatcgg agaatgcctt 4980tgacagcttt ctagtaacgg tggtggattc tgggaagctg ctggaccccc aggaattcac 5040actttcagga acccagagga agctggagct tagaggcctc ataactggca ttggctatga 5100ggttatggtc tctggcttca cccaagggca tcaaaccaag cccttgaggg ctgagattgt 5160tacagaagcc gaaccggaag ttgacaacct tctggtttca gatgccaccc cagacggttt 5220ccgtctgtcc tggacagctg atgaaggggt cttcgacaat tttgttctca aaatcagaga 5280taccaaaaag cagtctgagc cactggaaat aaccctactt gcccccgaac gtaccaggga 5340cttaacaggt ctcagagagg ctactgaata cgaaattgaa ctctatggaa taagcaaagg 5400aaggcgatcc cagacagtca gtgctatagc aacaacagcc atgggctccc caaaggaagt 5460cattttctca gacatcactg aaaattcggc tactgtcagc tggagggcac ccacggccca 5520agtggagagc ttccggatta cctatgtgcc cattacagga ggtacaccct ccatggtaac 5580tgtggacgga accaagactc agaccaggct ggtgaaactc atacctggcg tggagtacct 5640tgtcagcatc atcgccatga agggctttga ggaaagtgaa cctgtctcag ggtcattcac 5700cacagctctg gatggcccat ctggcctggt gacagccaac atcactgact cagaagcctt 5760ggccaggtgg cagccagcca ttgccactgt ggacagttat gtcatctcct acacaggcga 5820gaaagtgcca gaaattacac gcacggtgtc cgggaacaca gtggagtatg ctctgaccga 5880cctcgagcct gccacggaat acacactgag aatctttgca gagaaagggc cccagaagag 5940ctcaaccatc actgccaagt tcacaacaga cctcgattct ccaagagact tgactgctac 6000tgaggttcag tcggaaactg ccctccttac ctggcgaccc ccccgggcat cagtcaccgg 6060ttacctgctg gtctatgaat cagtggatgg cacagtcaag gaagtcattg tgggtccaga 6120taccacctcc tacagcctgg cagacctgag cccatccacc cactacacag ccaagatcca 6180ggcactcaat gggcccctga ggagcaatat gatccagacc atcttcacca caattggact 6240cctgtacccc ttccccaagg actgctccca agcaatgctg aatggagaca cgacctctgg 6300cctctacacc atttatctga atggtgataa ggctcaggcg ctggaagtct tctgtgacat 6360gacctctgat gggggtggat ggattgtgtt cctgagacgc aaaaacggac gcgagaactt 6420ctaccaaaac tggaaggcat atgctgctgg atttggggac cgcagagaag aattctggct 6480tgggctggac aacctgaaca aaatcacagc ccaggggcag tacgagctcc gggtggacct 6540gcgggaccat ggggagacag cctttgctgt ctatgacaag ttcagcgtgg gagatgccaa 6600gactcgctac aagctgaagg tggaggggta cagtgggaca gcaggtgact ccatggccta 6660ccacaatggc agatccttct ccacctttga caaggacaca gattcagcca tcaccaactg 6720tgctctgtcc tacaaagggg ctttctggta caggaactgt caccgtgtca acctgatggg 6780gagatatggg gacaataacc acagtcaggg cgttaactgg ttccactgga agggccacga 6840acactcaatc cagtttgctg agatgaagct gagaccaagc aacttcagaa atcttgaagg 6900caggcgcaaa cgggcataaa ttggagggac cactgggtga gagaggaata aggcggccca 6960gagcgaggaa aggattttac caaagcatca atacaaccag cccaaccatc ggtccacacc 7020tgggcatttg gtgagaatca aagctgacca tggatccctg gggccaacgg caacagcatg 7080ggcctcacct cctctgtgat ttctttcttt gcaccaaaga catcagtctc caacatgttt 7140ctgttttgtt gtttgattca gcaaaaatct cccagtgaca acatcgcaat agttttttac 7200ttctcttagg tggctctggg atgggagagg ggtaggatgt acaggggtag tttgttttag 7260aaccagccgt attttacatg aagctgtata attaattgtc attatttttg ttagcaaaga 7320ttaaatgtgt cattggaagc catccctttt tttacatttc atacaacaga aaccagaaaa 7380gcaatactgt ttccatttta aggatatgat taatattatt aatataataa tgatgatgat 7440gatgatgaaa actaaggatt tttcaagaga tctttctttc caaaacattt ctggacagta 7500cctgattgta tttttttttt aaataaaagc acaagtactt ttgaaaaaaa accggaattc 756016 2232 DNA Homo sapiens 16 cttccccttc tctgccctgc tccaggcacc aggctctttccccttcagtg tctcagagga 60 ggggacggca gcaccatgga cccccgcttg tccactgtccgccagacctg ctgctgcttc 120 aatgtccgca tcgcaaccac cgccctggcc atctaccatgtgatcatgag cgtcttgttg 180 ttcatcgagc actcagtaga ggtggcccat ggcaaggcgtcctgcaagct ctcccagatg 240 ggctacctca ggatcgctga cctgatctcc agcttcctgctcatcaccat gctcttcatc 300 atcagcctga gcctactgat cggcgtagtc aagaaccgggagaagtacct gctgcccttc 360 ctgtccctgc aaatcatgga ctatctcctg tgcctgctcaccctgctggg ctcctacatt 420 gagctgcccg cctacctcaa gttggcctcc cggagccgtgctagctcctc caagttcccc 480 ctgatgacgc tgcagctgct ggacttctgc ctgagcatcctgaccctctg cagctcctac 540 atggaagtgc ccacctatct caacttcaag tccatgaaccacatgaatta cctccccagc 600 caggaggata tgcctcataa ccagttcatc aagatgatgatcatcttttc catcgccttc 660 atcactgtcc ttatcttcaa ggtctacatg ttcaagtgcgtgtggcggtg ctacagattg 720 atcaagtgca tgaactcggt ggaggagaag agaaactccaagatgctcca gaaggtggtc 780 ctgccgtcct acgaggaagc cctgtctttg ccatcgaagaccccagaggg gggcccagca 840 ccacccccat actcagaggt gtgaccctcg ccaggccccagccccagtgc tgggaggggt 900 ggagctgcct cataatctgc ttttttgctt tggtggcccctgtggcctgg gtgggccctc 960 ccgcccctcc ctggcaggac aatctgcttg tgtctccctcgctggcctgc tcctcctgca 1020 gggcctgtga gctgctcaca actgggtcaa cgctttaggctgagtcactc ctcgggtctc 1080 tccataattc agcccaacaa tgcttggttt atttcaatcagctctgacac ttgtttagac 1140 gattggccat tctaaagttg gtgagtttgt caagcaactatcgacttgat cagttcagcc 1200 aagcaactga caaatcaaaa acccacttgt cagttcagtaaaataatttg gtcaaacaac 1260 agtctattgc attgatttat aaatagttgt cagttcacatagcaatttaa tcaagtaatc 1320 attaattagt taccccctat atataaatat atgtaatcaatttcttcaaa tagcttgctt 1380 acatgataat caattagcca accatgagtc atttagaatagtgataaata gaatacacag 1440 aatagtgatg aaattcaatt taaaaaatca cgttagcctccaaaccattt aattcaaatg 1500 aacccatcaa ctggatgcca actctggcga atgtaggacctctgagtggc tgtataattg 1560 ttaattcaaa tgaaattcat ttaaacagtt gacaaactgtcattcaacaa ttagctccag 1620 gaaataacag ttatttcatc ataaaacagt cccttcaaacacacaattgt tctgctgaag 1680 agttgtcatc aacaatccaa tgctcaccta ttcagttgctctgtggtcag tgtggctgca 1740 tagcagtgga ttccatgaaa ggagtcattt tagtgatgagctgccagtcc attcccaggc 1800 caggctgtcg ctggccatcc attcagtcga ttcagtcataggcgaatctg ttctgcccga 1860 ggcttgtggt caagcaaaaa ttcagccctg aaatcaggcacatctgttcg ttggactaaa 1920 cccacaggtt agttcagtca aagcaggcaa cccccttgtgggcactgacc ctgccactgg 1980 ggtcatggcg gttgtggcag ctggggaggt ttggccccaacagccctcct gtgcctgctt 2040 ccctgtgtgt cggggtcctc cagggagctg acccagaggtggaggccacg gaggcagggt 2100 ctctggggac tgtcgggggg tacagaggga gaaggctctgcaagagctcc ctggcaatac 2160 ccccttgtgt aattgctttg tgtgcgacag ggaggaagtttcaataaagc aacaacaagc 2220 ttcaaggaat tc 2232 17 2709 DNA Homo sapiens17 gggaatagca gaataggagc aagccagcac tagtcagcta actaagtgac tcaaccaagg 60ccttttttcc ttgttatctt tgcagatact tcattttctt agcgtttctg gagattacaa 120catcctgcgg ttccgtttct gggaacttta ctgatttatc tcccccctca cacaaataag 180cattgattcc tgcatttctg aagatctcaa gatctggact actgttgaaa aaatttccag 240tgaggctcac ttatgtctgt aaagatggga aaaaaataca agaacattgt tctactaaaa 300ggattagagg tcatcaatga ttatcatttt agaatggtta agtccttact gagcaacgat 360ttaaaactta atttaaaaat gagagaagag tatgacaaaa ttcagattgc tgacttgatg 420gaagaaaagt tccgaggtga tgctggtttg ggcaaactaa taaaaatttt cgaagatata 480ccaacgcttg aagacctggc tgaaactctt aaaaaagaaa agttaaaagt aaaaggacca 540gccctatcaa gaaagaggaa gaaggaagtg catgctactt cacctgcacc ctccacaagc 600agcactgtca aaactgaagg agcagaggca actcctggag ctcagaaaag aaaaaaatca 660accaaagaaa aggctggacc caaagggagt aaggtgtccg aggaacagac tcagcctccc 720tctcctgcag gagccggcat gtccacagcc atgggccgtt ccccatctcc caagacctca 780ttgtcagctc cacccaacag ttcttcaact gagaacccga aaacagtggc caaatgtcag 840gtaactccca gaagaaatgt tctccaaaaa cgcccagtga tagtgaaggt actgagtaca 900acaaagccat ttgaatatga gaccccagaa atggagaaaa aaataatgtt tcatgctaca 960gtggctacac agacacagtt cttccatgtg aaggttttaa acaccagctt gaaggagaaa 1020ttcaatggaa agaaaatcat catcatatca gattatttgg aatatgatag tctcctagag 1080gtcaatgaag aatctactgt atctgaagct ggtcctaacc aaacgtttga ggttccaaat 1140aaaatcatca acagagcaaa ggaaactctg aagattgata ttcttcacaa acaagcttca 1200ggaaatattg tatatggggt atttatgcta cataagaaaa cagtaaatca gaagaccaca 1260atctacgaaa ttcaggatga tagaggaaaa atggatgtag tggggacagg acaatgtcac 1320aatatcccct gtgaagaagg agataagctc cagcttttct gctttcgact tagaaaaaag 1380aaccagatgt caaaactgat ttcagaaatg catagtttta tccagataaa gaaaaaaaca 1440aacccgagaa acaatgaccc caagagcatg aagctacccc aggaacagcg tcagcttcca 1500tatccttcag aggccagcac aaccttccct gagagccatc ttcggactcc tcagatgcca 1560ccaacaactc catccagcag tttcttcacc aagaaaagtg aagacacaat ctccaaaatg 1620aatgacttca tgaggatgca gatactgaag gaagggagtc attttccagg accgttcatg 1680accagcatag gcccagctga gagccatccc cacactcctc agatgcctcc atcaacacca 1740agcagcagtt tcttaaccac gttgaaacca agactgaaga ctgaacctga agaagtttcc 1800atagaagaca gtgcccagag tgacctcaaa gaagtgatgg tgctgaacgc aacagaatca 1860tttgtatatg agcccaaaga gcagaagaaa atgtttcatg ccacagtggc aactgagaat 1920gaagtcttcc gagtgaaggt ttttaatatt gacctaaagg agaagttcac cccaaagaag 1980atcattgcca tagcaaatta tgtttgccgc aatgggttcc tggaggtata tcctttcaca 2040cttgtggctg atgtgaatgc tgaccgaaac atggagatcc caaaaggatt gattagaagt 2100gccagcgtaa ctcctaaaat caatcagctt tgctcacaaa ctaaaggaag ttttgtgaat 2160ggggtgtttg aggtacataa gaaaaatgta aggggtgaat tcacttatta tgaaatacaa 2220gataatacag ggaagatgga agtggtggtg catggacgac tgaacacaat caactgtgag 2280gaaggagata aactgaaact caccagcttt gaattggcac cgaaaagtgg gaataccggg 2340gagttgagat ctgtaattca tagtcacatc aaggtcatca agaccaggaa aaacaagaaa 2400gacatactca atcctgattc aagtatggaa acttcaccag actttttctt ctaaaatctg 2460gatgtcattg acgataatgt ttatggagat aaggtctaag tccctaaaaa aatgtacata 2520tacctggttg aaatacaaca ctatacatac acaccaccat atatactagc tgttaatcct 2580atggaatggg ggtattggga gtgctttttt aatttttcat agtttttttt taataaaatg 2640gcatattttg catctacaac ttctataata agaaaaaata aataaacatt atcttttttg 2700tgaaaaaaa 2709 18 1722 DNA Homo sapiens 18 gcgggcggtt cagccatgaggctggctgtg cttttctcgg gggccctgct ggggctactg 60 gcagcccagg ggacagggaatgactgtcct cacaaaaaat cagctacttt gctgccatcc 120 ttcacggtga cacccacggttacagagagc actggaacaa ccagccacag gactaccaag 180 agccacaaaa ccaccactcacaggacaacc accacaggca ccaccagcca cggacccacg 240 actgccactc acaaccccaccaccaccagc catggaaacg tcacagttca tccaacaagc 300 aatagcactg ccaccagccagggaccctca actgccactc acagtcctgc caccactagt 360 catggaaatg ccacggttcatccaacaagc aacagcactg ccaccagccc aggattcacc 420 agttctgccc acccagaaccacctccaccc tctccgagtc ctagcccaac ctccaaggag 480 accattggag actacacgtggaccaatggt tcccagccct gtgtccacct ccaagcccag 540 attcagattc gagtcatgtacacaacccag ggtggaggag aggcctgggg catctctgta 600 ctgaacccca acaaaaccaaggtccaggga agctgtgagg gtgcccatcc ccacctgctt 660 ctctcattcc cctatggacacctcagcttt ggattcatgc aggacctcca gcagaaggtt 720 gtctacctga gctacatggcggtggagtac aatgtgtcct tcccccacgc agcaaagtgg 780 acattctcgg ctcagaatgcatcccttcga gatctccaag cacccctggg gcagagcttc 840 agttgcagca actcgagcatcattctttca ccagctgtcc acctcgacct gctctccctg 900 aggctccagg ctgctcagctgccccacaca ggggtctttg ggcaaagttt ctcctgcccc 960 agtgaccggt ccatcttgctgcctctcatc atcggcctga tccttcttgg cctcctcgcc 1020 ctggtgctta ttgctttctgcatcatccgg agacgcccat ccgcctacca ggccctctga 1080 gcatttgctt caaaccccagggcactgagg gggtttgggg tgtggtgggg gggtaccctt 1140 atttcctcga cacgccgctggctcaaagac aatgttattt tccttccctt tcttgaagaa 1200 caaaaagaaa gccgggcatgacggctcatg cctgtaatcc cagcactttg ggaggctgag 1260 gcaggtggat cactggaggtcaggtctttg aggccagccc tagccaacat ggtgtaaaca 1320 ctgtctctac taaaaatacaattagccagg tgtggcggcg taatcccatg ctaacctgta 1380 atcccagcta cttgggaggctgaggcagag ctgcttgaac cctggaagtg gaggttgcag 1440 tgagcctgtc atcgctccactgagccaaga tcgctcccac tgcactccag cctgggcgac 1500 agagccagac tgtctcaaataaataaatat gagataatgc agtcgggaga agggagggag 1560 agaattttat taaatgtgacgaactgcccc cccccccccc cccagcagga gagcagcaaa 1620 atttatgtaa atctttgacggggttttcct tgctcctgcc aggattaaaa gtccatgagt 1680 ttcttgctca aaaaaaaaaaaaaaaaaaaa aaaaaaaaaa aa 1722 19 1522 DNA Homo sapiens 19 caggtgtggattccgccggt gaaggctgaa ggcagctacc ttaaagatgc cgggatccgc 60 agcgaagggctcggagttgt cagagaggat cgagagcttc gtggagaccc tgaagcgggg 120 tggtgggccgcgcagctccg aggaaatggc tcgggagacc ctagggttgc tgcgccagat 180 catcacggaccaccgctgga gcaacgcggg ggagctgatg gagctgatcc gcagagaggg 240 caggaggatgacggccgctc agccctccga gaccaccgtg ggcaacatgg tgcggagagt 300 gctcaagattatccgggagg agtatggcag actccatgga cgcagcgacg agagtgatca 360 gcaggagtccctgcacaaac tgttgacatc cggaggccta aacgaggatt tcagcttcca 420 ttatgcccaactccagtcca acatcattga ggcgattaat gagctgctag tggagctgga 480 agggacaatggagaacattg cagcccaggc tctggagcac attcactcca atgaggtgat 540 catgaccattggcttctccc gaacagtaga ggccttcctc aaagaggctg cccgaaagag 600 gaaattccatgtcattgtag cagagtgtgc tcctttctgc cagggtcatg aaatggctgt 660 gaatttgtccaaagcaggta ttgagacaac tgtcatgact gatgctgcca tttttgccgt 720 tatgtcaagagtcaacaagg tgatcattgg cacgaagacc atcctggcca atggggccct 780 gagagctgtgacaggaactc acactctggc actggcagca aaacaccatt ccaccccact 840 catcgtctgtgcacctatgt tcaaactttc tccacagttc cccaatgaag aagactcatt 900 tcataagtttgtggctcctg aagaagtcct gccattcaca gaaggggaca ttctggagaa 960 ggtcagcgtgcattgccctg tgtttgacta cgttccccca gagctcatta ccctctttat 1020 ctccaacattggtgggaatg caccttccta catctaccgc ctgatgagtg aactctacca 1080 tcctgatgatcatgttttat gaccgaccac acgtgtccta agcagattgc ttaggcagat 1140 acagaatgaagaggagactt gagtgttgct gctgaagcac atccttgcaa tgtgggagtg 1200 cacaggagtccacctaaaaa aaaaatcctt gatactgttg cctgcctttt tagtcacccc 1260 gtaacaagggcacacatcca gcactgtgtc ttgcctttca gatcttaaca gagcagcagg 1320 gcttaacttgttgattttgg agcctcttag tgacctggtt gcgtctgtgt caggaactta 1380 aactttctggttcagtagtg tgttaaacat aacactgaat accttactgg gatacagatt 1440 tttgctcagaaatggctatg acactttttc taggctctac caataaaagc cacttgaagg 1500 ttcaaaaaaaaaaaaaaaaa aa 1522 20 662 PRT Homo sapiens 20 Met Val Val Ser Glu ValAsp Ile Ala Lys Ala Asp Pro Ala Ala Ala 1 5 10 15 Ser His Pro Leu LeuLeu Asn Gly Asp Ala Thr Val Ala Gln Lys Asn 20 25 30 Pro Gly Ser Val AlaGlu Asn Asn Leu Cys Ser Gln Tyr Glu Glu Lys 35 40 45 Val Arg Pro Cys IleAsp Leu Ile Asp Ser Leu Arg Ala Leu Gly Val 50 55 60 Glu Gln Asp Leu AlaLeu Pro Ala Ile Ala Val Ile Gly Asp Gln Ser 65 70 75 80 Ser Gly Lys SerSer Val Leu Glu Ala Leu Ser Gly Val Ala Leu Pro 85 90 95 Arg Gly Ser GlyIle Val Thr Arg Cys Pro Leu Val Leu Lys Leu Lys 100 105 110 Lys Leu ValAsn Glu Asp Lys Trp Arg Gly Lys Val Ser Tyr Gln Asp 115 120 125 Tyr GluIle Glu Ile Ser Asp Ala Ser Glu Val Glu Lys Glu Ile Asn 130 135 140 LysAla Gln Asn Ala Ile Ala Gly Glu Gly Met Gly Ile Ser His Glu 145 150 155160 Leu Ile Thr Leu Glu Ile Ser Ser Arg Asp Val Pro Asp Leu Thr Leu 165170 175 Ile Asp Leu Pro Gly Ile Thr Arg Val Ala Val Gly Asn Gln Pro Ala180 185 190 Asp Ile Gly Tyr Lys Ile Lys Thr Leu Ile Lys Lys Tyr Ile GlnArg 195 200 205 Gln Glu Thr Ile Ser Leu Val Val Val Pro Ser Asn Val AspIle Ala 210 215 220 Thr Thr Glu Ala Leu Ser Met Ala Gln Glu Val Asp ProGlu Gly Asp 225 230 235 240 Arg Thr Ile Gly Ile Leu Thr Lys Pro Asp LeuVal Asp Lys Gly Thr 245 250 255 Glu Asp Lys Val Val Asp Val Val Arg AsnLeu Val Phe His Leu Lys 260 265 270 Lys Gly Tyr Met Ile Val Lys Cys ArgGly Gln Gln Glu Ile Gln Asp 275 280 285 Gln Leu Ser Leu Ser Glu Ala LeuGln Arg Glu Lys Ile Phe Phe Glu 290 295 300 Asn His Pro Tyr Phe Arg AspLeu Leu Glu Glu Gly Lys Ala Thr Val 305 310 315 320 Pro Cys Leu Ala GluLys Leu Thr Ser Glu Leu Ile Thr His Ile Cys 325 330 335 Lys Ser Leu ProLeu Leu Glu Asn Gln Ile Lys Glu Thr His Gln Arg 340 345 350 Ile Thr GluGlu Leu Gln Lys Tyr Gly Val Asp Ile Pro Glu Asp Glu 355 360 365 Asn GluLys Met Phe Phe Leu Ile Asp Lys Ile Asn Ala Phe Asn Gln 370 375 380 AspIle Thr Ala Leu Met Gln Gly Glu Glu Thr Val Gly Glu Glu Asp 385 390 395400 Ile Arg Leu Phe Thr Arg Leu Arg His Glu Phe His Lys Trp Ser Thr 405410 415 Ile Ile Glu Asn Asn Phe Gln Glu Gly His Lys Ile Leu Ser Arg Lys420 425 430 Ile Gln Lys Phe Glu Asn Gln Tyr Arg Gly Arg Glu Leu Pro GlyPhe 435 440 445 Val Asn Tyr Arg Thr Phe Glu Thr Ile Val Lys Gln Gln IleLys Ala 450 455 460 Leu Glu Glu Pro Ala Val Asp Met Leu His Thr Val ThrAsp Met Val 465 470 475 480 Arg Leu Ala Phe Thr Asp Val Ser Ile Lys AsnPhe Glu Glu Phe Phe 485 490 495 Asn Leu His Arg Thr Ala Lys Ser Lys IleGlu Asp Ile Arg Ala Glu 500 505 510 Gln Glu Arg Glu Gly Glu Lys Leu IleArg Leu His Phe Gln Met Glu 515 520 525 Gln Ile Val Tyr Cys Gln Asp GlnVal Tyr Arg Gly Ala Leu Gln Lys 530 535 540 Val Arg Glu Lys Glu Leu GluGlu Glu Lys Lys Lys Lys Ser Trp Asp 545 550 555 560 Phe Gly Ala Phe GlnSer Ser Ser Ala Thr Asp Ser Ser Met Glu Glu 565 570 575 Ile Phe Gln HisLeu Met Ala Tyr His Gln Glu Ala Ser Lys Arg Ile 580 585 590 Ser Ser HisIle Pro Leu Ile Ile Gln Phe Phe Met Leu Gln Thr Tyr 595 600 605 Gly GlnGln Leu Gln Lys Ala Met Leu Gln Leu Leu Gln Asp Lys Asp 610 615 620 ThrTyr Ser Trp Leu Leu Lys Glu Arg Ser Asp Thr Ser Asp Lys Arg 625 630 635640 Lys Phe Leu Lys Glu Arg Leu Ala Arg Leu Thr Gln Ala Arg Arg Arg 645650 655 Leu Ala Gln Phe Pro Gly 660 21 110 PRT Homo sapiens 21 Met SerMet Thr Asp Leu Leu Asn Ala Glu Asp Ile Lys Lys Ala Val 1 5 10 15 GlyAla Phe Ser Ala Thr Asp Ser Phe Asp His Lys Lys Phe Phe Gln 20 25 30 MetVal Gly Leu Lys Lys Lys Ser Ala Asp Asp Val Lys Lys Val Phe 35 40 45 HisMet Leu Asp Lys Asp Lys Ser Gly Phe Ile Glu Glu Asp Glu Leu 50 55 60 GlyPhe Ile Leu Lys Gly Phe Ser Pro Asp Ala Arg Asp Leu Ser Ala 65 70 75 80Lys Glu Thr Lys Met Leu Met Ala Ala Gly Asp Lys Asp Gly Asp Gly 85 90 95Lys Ile Gly Val Asp Glu Phe Ser Thr Leu Val Ala Glu Ser 100 105 110 221722 PRT Homo sapiens 22 Met Ala Gly Val Gly Pro Gly Gly Tyr Ala Ala GluPhe Val Pro Pro 1 5 10 15 Pro Glu Cys Pro Val Phe Glu Pro Ser Trp GluGlu Phe Thr Asp Pro 20 25 30 Leu Ser Phe Ile Gly Arg Ile Arg Pro Leu AlaGlu Lys Thr Gly Ile 35 40 45 Cys Lys Ile Arg Pro Pro Lys Asp Trp Gln ProPro Phe Ala Cys Glu 50 55 60 Val Lys Ser Phe Arg Phe Thr Pro Arg Val GlnArg Leu Asn Glu Leu 65 70 75 80 Glu Ala Met Thr Arg Val Arg Leu Asp PheLeu Asp Gln Leu Ala Lys 85 90 95 Phe Trp Glu Leu Gln Gly Ser Thr Leu LysIle Pro Val Val Glu Arg 100 105 110 Lys Ile Leu Asp Leu Tyr Ala Leu SerLys Ile Val Ala Ser Lys Gly 115 120 125 Gly Phe Glu Met Val Thr Lys GluLys Lys Trp Ser Lys Val Gly Ser 130 135 140 Arg Leu Gly Tyr Leu Pro GlyLys Gly Thr Gly Ser Leu Leu Lys Ser 145 150 155 160 His Tyr Glu Arg IleLeu Tyr Pro Tyr Glu Leu Phe Gln Ser Gly Val 165 170 175 Ser Leu Met GlyVal Gln Met Pro Asn Leu Asp Leu Lys Glu Lys Val 180 185 190 Glu Pro GluVal Leu Ser Thr Asp Thr Gln Thr Ser Pro Glu Pro Gly 195 200 205 Thr ArgMet Asn Ile Leu Pro Lys Arg Thr Arg Arg Val Lys Thr Gln 210 215 220 SerGlu Ser Gly Asp Val Ser Arg Asn Thr Glu Leu Lys Lys Leu Gln 225 230 235240 Ile Phe Gly Ala Gly Pro Lys Val Val Gly Leu Ala Met Gly Thr Lys 245250 255 Asp Lys Glu Asp Glu Val Thr Arg Arg Arg Lys Val Thr Asn Arg Ser260 265 270 Asp Ala Phe Asn Met Gln Met Arg Gln Arg Lys Gly Thr Leu SerVal 275 280 285 Asn Phe Val Asp Leu Tyr Val Cys Met Phe Cys Gly Arg GlyAsn Asn 290 295 300 Glu Asp Lys Leu Leu Leu Cys Asp Gly Cys Asp Asp SerTyr His Thr 305 310 315 320 Phe Cys Leu Ile Pro Pro Leu Pro Asp Val ProLys Gly Asp Trp Arg 325 330 335 Cys Pro Lys Cys Val Ala Glu Glu Cys SerLys Pro Arg Glu Ala Phe 340 345 350 Gly Phe Glu Gln Ala Val Arg Glu TyrThr Leu Gln Ser Phe Gly Glu 355 360 365 Met Ala Asp Asn Phe Lys Ser AspTyr Phe Asn Met Pro Val His Met 370 375 380 Val Pro Thr Glu Leu Val GluLys Glu Phe Trp Arg Leu Val Ser Ser 385 390 395 400 Ile Glu Glu Asp ValIle Val Glu Tyr Gly Ala Asp Ile Ser Ser Lys 405 410 415 Asp Phe Gly SerGly Phe Pro Val Lys Asp Gly Arg Arg Lys Ile Leu 420 425 430 Pro Glu GluGlu Glu Tyr Ala Leu Ser Gly Trp Asn Leu Asn Asn Met 435 440 445 Pro ValLeu Glu Gln Ser Val Leu Ala His Ile Asn Val Asp Ile Ser 450 455 460 GlyMet Lys Val Pro Trp Leu Tyr Val Gly Met Cys Phe Ser Ser Phe 465 470 475480 Cys Trp His Ile Glu Asp His Trp Ser Tyr Ser Ile Asn Tyr Leu His 485490 495 Trp Gly Glu Pro Lys Thr Trp Tyr Gly Val Pro Ser His Ala Ala Glu500 505 510 Gln Leu Glu Glu Val Met Arg Glu Leu Ala Pro Glu Leu Phe GluSer 515 520 525 Gln Pro Asp Leu Leu His Gln Leu Val Thr Ile Met Asn ProAsn Val 530 535 540 Leu Met Glu His Gly Val Pro Val Tyr Arg Thr Asn GlnCys Ala Gly 545 550 555 560 Glu Phe Val Val Thr Phe Pro Arg Ala Tyr HisSer Gly Phe Asn Gln 565 570 575 Gly Tyr Asn Phe Ala Glu Ala Val Asn PheCys Thr Ala Asp Trp Leu 580 585 590 Pro Ile Gly Arg Gln Cys Val Asn HisTyr Arg Arg Leu Arg Arg His 595 600 605 Cys Val Phe Ser His Glu Glu LeuIle Phe Lys Met Ala Ala Asp Pro 610 615 620 Glu Cys Leu Asp Val Gly LeuAla Ala Met Val Cys Lys Glu Leu Thr 625 630 635 640 Leu Met Thr Glu GluGlu Thr Arg Leu Arg Glu Ser Val Val Gln Met 645 650 655 Gly Val Leu MetSer Glu Glu Glu Val Phe Glu Leu Val Pro Asp Asp 660 665 670 Glu Arg GlnCys Ser Ala Cys Arg Thr Thr Cys Phe Leu Ser Ala Leu 675 680 685 Thr CysSer Cys Asn Pro Glu Arg Leu Val Cys Leu Tyr His Pro Thr 690 695 700 AspLeu Cys Pro Cys Pro Met Gln Lys Lys Cys Leu Arg Tyr Arg Tyr 705 710 715720 Pro Leu Glu Asp Leu Pro Ser Leu Leu Tyr Gly Val Lys Val Arg Ala 725730 735 Gln Ser Tyr Asp Thr Trp Val Ser Arg Val Thr Glu Ala Leu Ser Ala740 745 750 Asn Phe Asn His Lys Lys Asp Leu Ile Glu Leu Arg Val Met LeuGlu 755 760 765 Asp Ala Glu Asp Arg Lys Tyr Pro Glu Asn Asp Leu Phe ArgLys Leu 770 775 780 Arg Asp Ala Val Lys Glu Ala Glu Thr Cys Ala Ser ValAla Gln Leu 785 790 795 800 Leu Leu Ser Lys Lys Gln Lys His Arg Gln SerPro Asp Ser Gly Arg 805 810 815 Thr Arg Thr Lys Leu Thr Val Glu Glu LeuLys Ala Phe Val Gln Gln 820 825 830 Leu Phe Ser Leu Pro Cys Val Ile SerGln Ala Arg Gln Val Lys Asn 835 840 845 Leu Leu Asp Asp Val Glu Glu PheHis Glu Arg Ala Gln Glu Ala Met 850 855 860 Met Asp Glu Thr Pro Asp SerSer Lys Leu Gln Met Leu Ile Asp Met 865 870 875 880 Gly Ser Ser Leu TyrVal Glu Leu Pro Glu Leu Pro Arg Leu Lys Gln 885 890 895 Glu Leu Gln GlnAla Arg Trp Leu Asp Glu Val Arg Leu Thr Leu Ser 900 905 910 Asp Pro GlnGln Val Thr Leu Asp Val Met Lys Lys Leu Ile Asp Ser 915 920 925 Gly ValGly Leu Ala Pro His His Ala Val Glu Lys Ala Met Ala Glu 930 935 940 LeuGln Glu Leu Leu Thr Val Ser Glu Arg Trp Glu Glu Lys Ala Lys 945 950 955960 Val Cys Leu Gln Ala Arg Pro Arg His Ser Val Ala Ser Leu Glu Ser 965970 975 Ile Val Asn Glu Ala Lys Asn Ile Pro Ala Phe Leu Pro Asn Val Leu980 985 990 Ser Leu Lys Glu Ala Leu Gln Lys Ala Arg Glu Trp Thr Ala LysVal 995 1000 1005 Glu Ala Ile Gln Ser Gly Ser Asn Tyr Ala Tyr Leu GluGln Leu 1010 1015 1020 Glu Ser Leu Ser Ala Lys Gly Arg Pro Ile Pro ValArg Leu Glu 1025 1030 1035 Ala Leu Pro Gln Val Glu Ser Gln Val Ala AlaAla Arg Ala Trp 1040 1045 1050 Arg Glu Arg Thr Gly Arg Thr Phe Leu LysLys Asn Ser Ser His 1055 1060 1065 Thr Leu Leu Gln Val Leu Ser Pro ArgThr Asp Ile Gly Val Tyr 1070 1075 1080 Gly Ser Gly Lys Asn Arg Arg LysLys Val Lys Glu Leu Ile Glu 1085 1090 1095 Lys Glu Lys Glu Lys Asp LeuAsp Leu Glu Pro Leu Ser Asp Leu 1100 1105 1110 Glu Glu Gly Leu Glu GluThr Arg Asp Thr Ala Met Val Val Ala 1115 1120 1125 Val Phe Lys Glu ArgGlu Gln Lys Glu Ile Glu Ala Met His Ser 1130 1135 1140 Leu Arg Ala AlaAsn Leu Ala Lys Met Thr Met Val Asp Arg Ile 1145 1150 1155 Glu Glu ValLys Phe Cys Ile Cys Arg Lys Thr Ala Ser Gly Phe 1160 1165 1170 Met LeuGln Cys Glu Leu Cys Lys Asp Trp Phe His Asn Ser Cys 1175 1180 1185 ValPro Leu Pro Lys Ser Ser Ser Gln Lys Lys Gly Ser Ser Trp 1190 1195 1200Gln Ala Lys Glu Val Lys Phe Leu Cys Pro Leu Cys Met Arg Ser 1205 12101215 Arg Arg Pro Arg Leu Glu Thr Ile Leu Ser Leu Leu Val Ser Leu 12201225 1230 Gln Lys Leu Pro Val Arg Leu Pro Glu Gly Glu Ala Leu Gln Cys1235 1240 1245 Leu Thr Glu Arg Ala Met Ser Trp Gln Asp Arg Ala Arg GlnAla 1250 1255 1260 Leu Ala Thr Asp Glu Leu Ser Ser Ala Leu Ala Lys LeuSer Val 1265 1270 1275 Leu Ser Gln Arg Met Val Glu Gln Ala Ala Arg GluLys Thr Glu 1280 1285 1290 Lys Ile Ile Ser Ala Glu Leu Gln Lys Ala AlaAla Asn Pro Asp 1295 1300 1305 Leu Gln Gly His Leu Pro Ser Phe Gln GlnSer Ala Phe Asn Arg 1310 1315 1320 Val Val Ser Ser Val Ser Ser Ser ProArg Gln Thr Met Asp Tyr 1325 1330 1335 Asp Asp Glu Glu Thr Asp Ser AspGlu Asp Ile Arg Glu Thr Tyr 1340 1345 1350 Gly Tyr Asp Met Lys Asp ThrAla Ser Val Lys Ser Ser Ser Ser 1355 1360 1365 Leu Glu Pro Asn Leu PheCys Asp Glu Glu Ile Pro Ile Lys Ser 1370 1375 1380 Glu Glu Val Val ThrHis Met Trp Thr Ala Pro Ser Phe Cys Ala 1385 1390 1395 Glu His Ala TyrSer Ser Ala Ser Lys Ser Cys Ser Gln Val Phe 1400 1405 1410 Phe Gly LysGly Ser Ser Thr Pro Arg Lys Gln Pro Arg Lys Ser 1415 1420 1425 Pro LeuVal Pro Arg Ser Leu Glu Pro Pro Val Leu Glu Leu Ser 1430 1435 1440 ProGly Ala Lys Ala Gln Leu Glu Glu Leu Met Met Val Gly Asp 1445 1450 1455Leu Leu Glu Val Ser Leu Asp Glu Thr Gln His Ile Trp Arg Ile 1460 14651470 Leu Gln Ala Thr His Pro Pro Ser Glu Asp Arg Phe Leu His Ile 14751480 1485 Met Glu Asp Asp Ser Met Glu Glu Lys Pro Leu Lys Val Lys Gly1490 1495 1500 Lys Asp Ser Ser Glu Lys Lys Arg Lys Arg Lys Leu Glu LysVal 1505 1510 1515 Glu Gln Leu Phe Gly Glu Gly Lys Gln Lys Ser Lys GluLeu Lys 1520 1525 1530 Lys Met Asp Lys Pro Arg Lys Lys Lys Leu Lys LeuGly Ala Asp 1535 1540 1545 Lys Ser Lys Lys Leu Asn Lys Leu Ala Lys LysLeu Ala Lys Glu 1550 1555 1560 Glu Glu Arg Lys Lys Lys Lys Glu Lys AlaAla Ala Ala Lys Val 1565 1570 1575 Glu Leu Val Lys Glu Ser Thr Glu LysLys Arg Glu Lys Lys Val 1580 1585 1590 Leu Asp Ile Pro Ser Lys Tyr AspTrp Ser Gly Ala Glu Glu Ser 1595 1600 1605 Asp Asp Glu Asn Ala Val CysAla Glu Pro Asp Cys Gln Arg Pro 1610 1615 1620 Cys Lys Asp Lys Gly ValVal Phe Val Thr Lys Lys Arg Glu Ile 1625 1630 1635 Lys Asn Ile Ser PheLys Ser Val Leu Cys Asp Cys Phe Ser Lys 1640 1645 1650 Lys Val Asp TrpVal Gln Cys Asp Gly Gly Cys Asp Glu Trp Phe 1655 1660 1665 His Arg ValCys Val Gly Val Ser Pro Glu Met Ala Glu Asn Glu 1670 1675 1680 Asp TyrIle Cys Ile Asn Cys Ala Lys Lys Gln Gly Pro Val Ser 1685 1690 1695 ProGly Pro Ala Pro Pro Pro Ser Phe Ile Met Ser Tyr Lys Leu 1700 1705 1710Pro Met Glu Asp Leu Lys Glu Thr Ser 1715 1720 23 373 PRT Homo sapiens 23Met Gly Ser Gln Val Ser Val Glu Ser Gly Ala Leu His Val Val Ile 1 5 1015 Val Gly Gly Gly Phe Gly Gly Ile Ala Ala Ala Ser Gln Leu Gln Ala 20 2530 Leu Asn Val Pro Phe Met Leu Val Asp Met Lys Asp Ser Phe His His 35 4045 Asn Val Ala Ala Leu Arg Ala Ser Val Glu Thr Gly Phe Ala Lys Lys 50 5560 Thr Phe Ile Ser Tyr Ser Val Thr Phe Lys Asp Asn Phe Arg Gln Gly 65 7075 80 Leu Val Val Gly Ile Asp Leu Lys Asn Gln Met Val Leu Leu Gln Gly 8590 95 Gly Glu Ala Leu Pro Phe Ser His Leu Ile Leu Ala Thr Gly Ser Thr100 105 110 Gly Pro Phe Pro Gly Lys Phe Asn Glu Val Ser Ser Gln Gln AlaAla 115 120 125 Ile Gln Ala Tyr Glu Asp Met Val Arg Gln Val Gln Arg SerArg Phe 130 135 140 Ile Val Val Val Gly Gly Gly Ser Ala Gly Val Glu MetAla Ala Glu 145 150 155 160 Ile Lys Thr Glu Tyr Pro Glu Lys Glu Val ThrLeu Ile His Ser Gln 165 170 175 Val Ala Leu Ala Asp Lys Glu Leu Leu ProSer Val Arg Gln Glu Val 180 185 190 Lys Glu Ile Leu Leu Arg Lys Gly ValGln Leu Leu Leu Ser Glu Arg 195 200 205 Val Ser Asn Leu Glu Glu Leu ProLeu Asn Glu Tyr Arg Glu Tyr Ile 210 215 220 Lys Val Gln Thr Asp Lys GlyThr Glu Val Ala Thr Asn Leu Val Ile 225 230 235 240 Leu Cys Thr Gly IleLys Ile Asn Ser Ser Ala Tyr Arg Lys Ala Phe 245 250 255 Glu Ser Arg LeuAla Ser Ser Gly Ala Leu Arg Val Asn Glu His Leu 260 265 270 Gln Val GluGly His Ser Asn Val Tyr Ala Ile Gly Asp Cys Ala Asp 275 280 285 Val ArgThr Pro Lys Met Ala Tyr Leu Ala Gly Leu His Ala Asn Ile 290 295 300 AlaVal Ala Asn Ile Val Asn Ser Val Lys Gln Arg Pro Leu Gln Ala 305 310 315320 Tyr Lys Pro Gly Ala Leu Thr Phe Leu Leu Ser Met Gly Arg Asn Asp 325330 335 Gly Val Gly Gln Ile Ser Gly Phe Tyr Val Gly Arg Leu Met Val Arg340 345 350 Leu Thr Lys Ser Arg Asp Leu Phe Val Ser Thr Ser Trp Lys ThrMet 355 360 365 Arg Gln Ser Pro Pro 370 24 209 PRT Homo sapiens 24 MetAla Ser Met Gly Leu Gln Val Met Gly Ile Ala Leu Ala Val Leu 1 5 10 15Gly Trp Leu Ala Val Met Leu Cys Cys Ala Leu Pro Met Trp Arg Val 20 25 30Thr Ala Phe Ile Gly Ser Asn Ile Val Thr Ser Gln Thr Ile Trp Glu 35 40 45Gly Leu Trp Met Asn Cys Val Val Gln Ser Thr Gly Gln Met Gln Cys 50 55 60Lys Val Tyr Asp Ser Leu Leu Ala Leu Pro Gln Asp Leu Gln Ala Ala 65 70 7580 Arg Ala Leu Val Ile Ile Ser Ile Ile Val Ala Ala Leu Gly Val Leu 85 9095 Leu Ser Val Val Gly Gly Lys Cys Thr Asn Cys Leu Glu Asp Glu Ser 100105 110 Ala Lys Ala Lys Thr Met Ile Val Ala Gly Val Val Phe Leu Leu Ala115 120 125 Gly Leu Met Val Ile Val Pro Val Ser Trp Thr Ala His Asn IleIle 130 135 140 Gln Asp Phe Tyr Asn Pro Leu Val Ala Ser Gly Gln Lys ArgGlu Met 145 150 155 160 Gly Ala Ser Leu Tyr Val Gly Trp Ala Ala Ser GlyLeu Leu Leu Leu 165 170 175 Gly Gly Gly Leu Leu Cys Cys Asn Cys Pro ProArg Thr Asp Lys Pro 180 185 190 Tyr Ser Ala Lys Tyr Ser Ala Ala Arg SerAla Ala Ala Ser Asn Tyr 195 200 205 Val 25 422 PRT Homo sapiens 25 MetAsn Ser Gly His Ser Phe Ser Gln Thr Pro Ser Ala Ser Phe His 1 5 10 15Gly Ala Gly Gly Gly Trp Gly Arg Pro Arg Ser Phe Pro Arg Ala Pro 20 25 30Thr Val His Gly Gly Ala Gly Gly Ala Arg Ile Ser Leu Ser Phe Thr 35 40 45Thr Arg Ser Cys Pro Pro Pro Gly Gly Ser Trp Gly Ser Gly Arg Ser 50 55 60Ser Pro Leu Leu Gly Gly Asn Gly Lys Ala Thr Met Gln Asn Leu Asn 65 70 7580 Asp Arg Leu Ala Ser Tyr Leu Glu Lys Val Arg Ala Leu Glu Glu Ala 85 9095 Asn Met Lys Leu Glu Ser Arg Ile Leu Lys Trp His Gln Gln Arg Asp 100105 110 Pro Gly Ser Lys Lys Asp Tyr Ser Gln Tyr Glu Glu Asn Ile Thr His115 120 125 Leu Gln Glu Gln Ile Val Asp Gly Lys Met Thr Asn Ala Gln IleIle 130 135 140 Leu Leu Ile Asp Asn Ala Arg Met Ala Val Asp Asp Phe AsnLeu Lys 145 150 155 160 Tyr Glu Asn Glu His Ser Phe Lys Lys Asp Leu GluIle Glu Val Glu 165 170 175 Gly Leu Arg Arg Thr Leu Asp Asn Leu Thr IleVal Thr Thr Asp Leu 180 185 190 Glu Gln Glu Val Glu Gly Met Arg Lys GluLeu Ile Leu Met Lys Lys 195 200 205 His His Glu Gln Glu Met Glu Lys HisHis Val Pro Ser Asp Phe Asn 210 215 220 Val Asn Val Lys Val Asp Thr GlyPro Arg Glu Asp Leu Ile Lys Val 225 230 235 240 Leu Glu Asp Met Arg GlnGlu Tyr Glu Leu Ile Ile Lys Lys Lys His 245 250 255 Arg Asp Leu Asp ThrTrp Tyr Lys Glu Gln Ser Ala Ala Met Ser Gln 260 265 270 Glu Ala Ala SerPro Ala Thr Val Gln Ser Arg Gln Gly Asp Ile His 275 280 285 Glu Leu LysArg Thr Phe Gln Ala Leu Glu Ile Asp Leu Gln Thr Gln 290 295 300 Tyr SerThr Lys Ser Ala Leu Glu Asn Met Leu Ser Glu Thr Gln Ser 305 310 315 320Arg Tyr Ser Cys Lys Leu Gln Asp Met Gln Glu Ile Ile Ser His Tyr 325 330335 Glu Glu Glu Leu Thr Gln Leu Arg His Glu Leu Glu Arg Gln Asn Asn 340345 350 Glu Tyr Gln Val Leu Leu Gly Ile Lys Thr His Leu Glu Lys Glu Ile355 360 365 Thr Thr Tyr Arg Arg Leu Leu Glu Gly Glu Ser Glu Gly Thr ArgGlu 370 375 380 Glu Ser Lys Ser Ser Met Lys Val Ser Ala Thr Pro Lys IleLys Ala 385 390 395 400 Ile Thr Gln Glu Thr Ile Asn Gly Arg Leu Val LeuCys Gln Val Asn 405 410 415 Glu Ile Gln Lys His Ala 420 26 541 PRT Homosapiens 26 Met Val Ala Asp Pro Pro Arg Asp Ser Lys Gly Leu Ala Ala AlaGlu 1 5 10 15 Pro Thr Ala Asn Gly Gly Leu Ala Leu Ala Ser Ile Glu AspGln Gly 20 25 30 Ala Ala Ala Gly Gly Tyr Cys Gly Ser Arg Asp Gln Val ArgArg Cys 35 40 45 Leu Arg Ala Asn Leu Leu Val Leu Leu Thr Val Val Ala ValVal Ala 50 55 60 Gly Val Ala Leu Gly Leu Gly Val Ser Gly Ala Gly Gly AlaLeu Ala 65 70 75 80 Leu Gly Pro Glu Arg Leu Ser Ala Phe Val Phe Pro GlyGlu Leu Leu 85 90 95 Leu Arg Leu Leu Arg Met Ile Ile Leu Pro Leu Val ValCys Ser Leu 100 105 110 Ile Gly Gly Ala Ala Ser Leu Asp Pro Gly Ala LeuGly Arg Leu Gly 115 120 125 Ala Trp Ala Leu Leu Phe Phe Leu Val Thr ThrLeu Leu Ala Ser Ala 130 135 140 Leu Gly Val Gly Leu Ala Leu Ala Leu GlnPro Gly Ala Ala Ser Ala 145 150 155 160 Ala Ile Asn Ala Ser Val Gly AlaAla Gly Ser Ala Glu Asn Ala Pro 165 170 175 Ser Lys Glu Val Leu Asp SerPhe Leu Asp Leu Ala Arg Asn Ile Phe 180 185 190 Pro Ser Asn Leu Val SerAla Ala Phe Arg Ser Tyr Ser Thr Thr Tyr 195 200 205 Glu Glu Arg Asn IleThr Gly Thr Arg Val Lys Val Pro Val Gly Gln 210 215 220 Glu Val Glu GlyMet Asn Ile Leu Gly Leu Val Val Phe Ala Ile Val 225 230 235 240 Phe GlyVal Ala Leu Arg Lys Leu Gly Pro Glu Gly Glu Leu Leu Ile 245 250 255 ArgPhe Phe Asn Ser Phe Asn Glu Ala Thr Met Val Leu Val Ser Trp 260 265 270Ile Met Trp Tyr Ala Pro Val Gly Ile Met Phe Leu Val Ala Gly Lys 275 280285 Ile Val Glu Met Glu Asp Val Gly Leu Leu Phe Ala Arg Leu Gly Lys 290295 300 Tyr Ile Leu Cys Cys Leu Leu Gly His Ala Ile His Gly Leu Leu Val305 310 315 320 Leu Pro Leu Ile Tyr Phe Leu Phe Thr Arg Lys Asn Pro TyrArg Phe 325 330 335 Leu Trp Gly Ile Val Thr Pro Leu Ala Thr Ala Phe GlyThr Ser Ser 340 345 350 Ser Ser Ala Thr Leu Pro Leu Met Met Lys Cys ValGlu Glu Asn Asn 355 360 365 Gly Val Ala Lys His Ile Ser Arg Phe Ile LeuPro Ile Gly Ala Thr 370 375 380 Val Asn Met Asp Gly Ala Ala Leu Phe GlnCys Val Ala Ala Val Phe 385 390 395 400 Ile Ala Gln Leu Ser Gln Gln SerLeu Asp Phe Val Lys Ile Ile Thr 405 410 415 Ile Leu Val Thr Ala Thr AlaSer Ser Val Gly Ala Ala Gly Ile Pro 420 425 430 Ala Gly Gly Val Leu ThrLeu Ala Ile Ile Leu Glu Ala Val Asn Leu 435 440 445 Pro Val Asp His IleSer Leu Ile Leu Ala Val Asp Trp Leu Val Asp 450 455 460 Arg Ser Cys ThrVal Leu Asn Val Glu Gly Asp Ala Leu Gly Ala Gly 465 470 475 480 Leu LeuGln Asn Tyr Val Asp Arg Thr Glu Ser Arg Ser Thr Glu Pro 485 490 495 GluLeu Ile Gln Val Lys Ser Glu Leu Pro Leu Asp Pro Leu Pro Val 500 505 510Pro Thr Glu Glu Gly Asn Pro Leu Leu Lys His Tyr Arg Gly Pro Ala 515 520525 Gly Asp Ala Thr Val Ala Ser Glu Lys Glu Ser Val Met 530 535 540 27472 PRT Homo sapiens 27 Met Ala Gly Gly Glu Ala Gly Val Thr Leu Gly GlnPro His Leu Ser 1 5 10 15 Arg Gln Asp Leu Thr Thr Leu Asp Val Thr LysLeu Thr Pro Leu Ser 20 25 30 His Glu Val Ile Ser Arg Gln Ala Thr Ile AsnIle Gly Thr Ile Gly 35 40 45 His Val Ala His Gly Lys Ser Thr Val Val LysAla Ile Ser Gly Val 50 55 60 His Thr Val Arg Phe Lys Asn Glu Leu Glu ArgAsn Ile Thr Ile Lys 65 70 75 80 Leu Gly Tyr Ala Asn Ala Lys Ile Tyr LysLeu Asp Asp Pro Ser Cys 85 90 95 Pro Arg Pro Glu Cys Tyr Arg Ser Cys GlySer Ser Thr Pro Asp Glu 100 105 110 Phe Pro Thr Asp Ile Pro Gly Thr LysGly Asn Phe Lys Leu Val Arg 115 120 125 His Val Ser Phe Val Asp Cys ProGly His Asp Ile Leu Met Ala Thr 130 135 140 Met Leu Asn Gly Ala Ala ValMet Asp Ala Ala Leu Leu Leu Ile Ala 145 150 155 160 Gly Asn Glu Ser CysPro Gln Pro Gln Thr Ser Glu His Leu Ala Ala 165 170 175 Ile Glu Ile MetLys Leu Lys His Ile Leu Ile Leu Gln Asn Lys Ile 180 185 190 Asp Leu ValLys Glu Ser Gln Ala Lys Glu Gln Tyr Glu Gln Ile Leu 195 200 205 Ala PheVal Gln Gly Thr Val Ala Glu Gly Ala Pro Ile Ile Pro Ile 210 215 220 SerAla Gln Leu Lys Tyr Asn Ile Glu Val Val Cys Glu Tyr Ile Val 225 230 235240 Lys Lys Ile Pro Val Pro Pro Arg Asp Phe Thr Ser Glu Pro Arg Leu 245250 255 Ile Val Ile Arg Ser Phe Asp Val Asn Lys Pro Gly Cys Glu Val Asp260 265 270 Asp Leu Lys Gly Gly Val Ala Gly Gly Ser Ile Leu Lys Gly ValLeu 275 280 285 Lys Val Gly Gln Glu Ile Glu Val Arg Pro Gly Ile Val SerLys Asp 290 295 300 Ser Glu Gly Lys Leu Met Cys Lys Pro Ile Phe Ser LysIle Val Ser 305 310 315 320 Leu Phe Ala Glu His Asn Asp Leu Gln Tyr AlaAla Pro Gly Gly Leu 325 330 335 Ile Gly Val Gly Thr Lys Ile Asp Pro ThrLeu Cys Arg Ala Asp Arg 340 345 350 Met Val Gly Gln Val Leu Gly Ala ValGly Ala Leu Pro Glu Ile Phe 355 360 365 Thr Glu Leu Glu Ile Ser Tyr PheLeu Leu Arg Arg Leu Leu Gly Val 370 375 380 Arg Thr Glu Gly Asp Lys LysAla Ala Lys Val Gln Lys Leu Ser Lys 385 390 395 400 Asn Glu Val Leu MetVal Asn Ile Gly Ser Leu Ser Thr Gly Gly Arg 405 410 415 Val Ser Ala ValLys Ala Asp Leu Gly Lys Ile Val Leu Thr Asn Pro 420 425 430 Val Cys ThrGlu Val Gly Glu Lys Ile Ala Leu Ser Arg Arg Val Glu 435 440 445 Lys HisTrp Arg Leu Ile Gly Trp Gly Gln Ile Arg Arg Gly Val Thr 450 455 460 IleLys Pro Thr Val Asp Asp Asp 465 470 28 669 PRT Homo sapiens 28 Met GluGly Asn Lys Leu Glu Glu Gln Asp Ser Ser Pro Pro Gln Ser 1 5 10 15 ThrPro Gly Leu Met Lys Gly Asn Lys Arg Glu Glu Gln Gly Leu Gly 20 25 30 ProGlu Pro Ala Ala Pro Gln Gln Pro Thr Ala Glu Glu Glu Ala Leu 35 40 45 IleGlu Phe His Arg Ser Tyr Arg Glu Leu Phe Glu Phe Phe Cys Asn 50 55 60 AsnThr Thr Ile His Gly Ala Ile Arg Leu Val Cys Ser Gln His Asn 65 70 75 80Arg Met Lys Thr Ala Phe Trp Ala Val Leu Trp Leu Cys Thr Phe Gly 85 90 95Met Met Tyr Trp Gln Phe Gly Leu Leu Phe Gly Glu Tyr Phe Ser Tyr 100 105110 Pro Val Ser Leu Asn Ile Asn Leu Asn Ser Asp Lys Leu Val Phe Pro 115120 125 Ala Val Thr Ile Cys Thr Leu Asn Pro Tyr Arg Tyr Pro Glu Ile Lys130 135 140 Glu Glu Leu Glu Glu Leu Asp Arg Ile Thr Glu Gln Thr Leu PheAsp 145 150 155 160 Leu Tyr Lys Tyr Ser Ser Phe Thr Thr Leu Val Ala GlySer Arg Ser 165 170 175 Arg Arg Asp Leu Arg Gly Thr Leu Pro His Pro LeuGln Arg Leu Arg 180 185 190 Val Pro Pro Pro Pro His Gly Ala Arg Arg AlaArg Ser Val Ala Ser 195 200 205 Ser Leu Arg Asp Asn Asn Pro Gln Val AspTrp Lys Asp Trp Lys Ile 210 215 220 Gly Phe Gln Leu Cys Asn Gln Asn LysSer Asp Cys Phe Tyr Gln Thr 225 230 235 240 Tyr Ser Ser Gly Val Asp AlaVal Arg Glu Trp Tyr Arg Phe His Tyr 245 250 255 Ile Asn Ile Leu Ser ArgLeu Pro Glu Thr Leu Pro Ser Leu Glu Glu 260 265 270 Asp Thr Leu Gly AsnPhe Ile Phe Ala Cys Arg Phe Asn Gln Val Ser 275 280 285 Cys Asn Gln AlaAsn Tyr Ser His Phe His His Pro Met Tyr Gly Asn 290 295 300 Cys Tyr ThrPhe Asn Asp Lys Asn Asn Ser Asn Leu Trp Met Ser Ser 305 310 315 320 MetPro Gly Ile Asn Asn Gly Leu Ser Leu Met Leu Arg Ala Glu Gln 325 330 335Asn Asp Phe Ile Pro Leu Leu Ser Thr Val Thr Gly Ala Arg Val Met 340 345350 Val His Gly Gln Asp Glu Pro Ala Phe Met Asp Asp Gly Gly Phe Asn 355360 365 Leu Arg Pro Gly Val Glu Thr Ser Ile Ser Met Arg Lys Glu Thr Leu370 375 380 Asp Arg Leu Gly Gly Asp Tyr Gly Asp Cys Thr Lys Asn Gly SerAsp 385 390 395 400 Val Pro Val Glu Asn Leu Tyr Pro Ser Lys Tyr Thr GlnGln Val Cys 405 410 415 Ile His Ser Cys Phe Gln Glu Ser Met Ile Lys GluCys Gly Cys Ala 420 425 430 Tyr Ile Phe Tyr Pro Arg Pro Gln Asn Val GluTyr Cys Asp Tyr Arg 435 440 445 Lys His Ser Ser Trp Gly Tyr Cys Tyr TyrLys Leu Gln Val Asp Phe 450 455 460 Ser Ser Asp His Leu Gly Cys Phe ThrLys Cys Arg Lys Pro Cys Ser 465 470 475 480 Val Thr Ser Tyr Gln Leu SerAla Gly Tyr Ser Arg Trp Pro Ser Val 485 490 495 Thr Ser Gln Glu Trp ValPhe Gln Met Leu Ser Arg Gln Asn Asn Tyr 500 505 510 Thr Val Asn Asn LysArg Asn Gly Val Ala Lys Val Asn Ile Phe Phe 515 520 525 Lys Glu Leu AsnTyr Lys Thr Asn Ser Glu Ser Pro Ser Val Thr Met 530 535 540 Val Thr LeuLeu Ser Asn Leu Gly Ser Gln Trp Ser Leu Trp Phe Gly 545 550 555 560 SerSer Val Leu Ser Val Val Glu Met Ala Glu Leu Val Phe Asp Leu 565 570 575Leu Val Ile Met Phe Leu Met Leu Leu Arg Arg Phe Arg Ser Arg Tyr 580 585590 Trp Ser Pro Gly Arg Gly Gly Arg Gly Ala Gln Glu Val Ala Ser Thr 595600 605 Leu Ala Ser Ser Pro Pro Ser His Phe Cys Pro His Pro Met Ser Leu610 615 620 Ser Leu Ser Gln Pro Gly Pro Ala Pro Ser Pro Ala Leu Thr AlaPro 625 630 635 640 Pro Pro Ala Tyr Ala Thr Leu Gly Pro Arg Pro Ser ProGly Gly Ser 645 650 655 Ala Gly Ala Ser Ser Ser Thr Cys Pro Leu Gly GlyPro 660 665 29 575 PRT Homo sapiens 29 Met Leu Gly Val Leu Val Leu GlyAla Leu Ala Leu Ala Gly Leu Gly 1 5 10 15 Phe Pro Ala Pro Ala Glu ProGln Pro Gly Gly Ser Gln Cys Val Glu 20 25 30 His Asp Cys Phe Ala Leu TyrPro Gly Pro Ala Thr Phe Leu Asn Ala 35 40 45 Ser Gln Ile Cys Asp Gly LeuArg Gly His Leu Met Thr Val Arg Ser 50 55 60 Ser Val Ala Ala Asp Val IleSer Leu Leu Leu Asn Gly Asp Gly Gly 65 70 75 80 Val Gly Arg Arg Arg LeuTrp Ile Gly Leu Gln Leu Pro Pro Gly Cys 85 90 95 Gly Asp Pro Lys Arg LeuGly Pro Leu Arg Gly Phe Gln Trp Val Thr 100 105 110 Gly Asp Asn Asn ThrSer Tyr Ser Arg Trp Ala Arg Leu Asp Leu Asn 115 120 125 Gly Ala Pro LeuCys Gly Pro Leu Cys Val Ala Val Ser Ala Ala Glu 130 135 140 Ala Thr ValPro Ser Glu Pro Ile Trp Glu Glu Gln Gln Cys Glu Val 145 150 155 160 LysAla Asp Gly Phe Leu Cys Glu Phe His Phe Pro Ala Thr Cys Arg 165 170 175Pro Leu Ala Val Glu Pro Gly Ala Ala Ala Ala Ala Val Ser Ile Thr 180 185190 Tyr Gly Thr Pro Phe Ala Ala Arg Gly Ala Asp Phe Gln Ala Leu Pro 195200 205 Val Gly Ser Ser Ala Ala Val Ala Pro Leu Gly Leu Gln Leu Met Cys210 215 220 Thr Ala Pro Pro Gly Ala Val Gln Gly His Trp Ala Arg Glu AlaPro 225 230 235 240 Gly Ala Trp Asp Cys Ser Val Glu Asn Gly Gly Cys GluHis Ala Cys 245 250 255 Asn Ala Ile Pro Gly Ala Pro Arg Cys Gln Cys ProAla Gly Ala Ala 260 265 270 Leu Gln Ala Asp Gly Arg Ser Cys Thr Ala SerAla Thr Gln Ser Cys 275 280 285 Asn Asp Leu Cys Glu His Phe Cys Val ProAsn Pro Asp Gln Pro Gly 290 295 300 Ser Tyr Ser Cys Met Cys Glu Thr GlyTyr Arg Leu Ala Ala Asp Gln 305 310 315 320 His Arg Cys Glu Asp Val AspAsp Cys Ile Leu Glu Pro Ser Pro Cys 325 330 335 Pro Gln Arg Cys Val AsnThr Gln Gly Gly Phe Glu Cys His Cys Tyr 340 345 350 Pro Asn Tyr Asp LeuVal Asp Gly Glu Cys Val Glu Pro Val Asp Pro 355 360 365 Cys Phe Arg AlaAsn Cys Glu Tyr Gln Cys Gln Pro Leu Asn Gln Thr 370 375 380 Ser Tyr LeuCys Val Cys Ala Glu Gly Phe Ala Pro Ile Pro His Glu 385 390 395 400 ProHis Arg Cys Gln Met Phe Cys Asn Gln Thr Ala Cys Pro Ala Asp 405 410 415Cys Asp Pro Asn Thr Gln Ala Ser Cys Glu Cys Pro Glu Gly Tyr Ile 420 425430 Leu Asp Asp Gly Phe Ile Cys Thr Asp Ile Asp Glu Cys Glu Asn Gly 435440 445 Gly Phe Cys Ser Gly Val Cys His Asn Leu Pro Gly Thr Phe Glu Cys450 455 460 Ile Cys Gly Pro Asp Ser Ala Leu Ala Arg His Ile Gly Thr AspCys 465 470 475 480 Asp Ser Gly Lys Val Asp Gly Gly Asp Ser Gly Ser GlyGlu Pro Pro 485 490 495 Pro Ser Pro Thr Pro Gly Ser Thr Leu Thr Pro ProAla Val Gly Leu 500 505 510 Val His Ser Gly Leu Leu Ile Gly Ile Ser IleAla Ser Leu Cys Leu 515 520 525 Val Val Ala Leu Leu Ala Leu Leu Cys HisLeu Arg Lys Lys Gln Gly 530 535 540 Ala Ala Arg Ala Lys Met Glu Tyr LysCys Ala Ala Pro Ser Lys Glu 545 550 555 560 Val Val Leu Gln His Val ArgThr Glu Arg Thr Pro Gln Arg Leu 565 570 575 30 604 PRT Homo sapiens 30Met Leu Ala Arg Ala Leu Leu Leu Cys Ala Val Leu Ala Leu Ser His 1 5 1015 Thr Ala Asn Pro Cys Cys Ser His Pro Cys Gln Asn Arg Gly Val Cys 20 2530 Met Ser Val Gly Phe Asp Gln Tyr Lys Cys Asp Cys Thr Arg Thr Gly 35 4045 Phe Tyr Gly Glu Asn Cys Ser Thr Pro Glu Phe Leu Thr Arg Ile Lys 50 5560 Leu Phe Leu Lys Pro Thr Pro Asn Thr Val His Tyr Ile Leu Thr His 65 7075 80 Phe Lys Gly Phe Trp Asn Val Val Asn Asn Ile Pro Phe Leu Arg Asn 8590 95 Ala Ile Met Ser Tyr Val Leu Thr Ser Arg Ser His Leu Ile Asp Ser100 105 110 Pro Pro Thr Tyr Asn Ala Asp Tyr Gly Tyr Lys Ser Trp Glu AlaPhe 115 120 125 Ser Asn Leu Ser Tyr Tyr Thr Arg Ala Leu Pro Pro Val ProAsp Asp 130 135 140 Cys Pro Thr Pro Leu Gly Val Lys Gly Lys Lys Gln LeuPro Asp Ser 145 150 155 160 Asn Glu Ile Val Glu Lys Leu Leu Leu Arg ArgLys Phe Ile Pro Asp 165 170 175 Pro Gln Gly Ser Asn Met Met Phe Ala PhePhe Ala Gln His Phe Thr 180 185 190 His Gln Phe Phe Lys Thr Asp His LysArg Gly Pro Ala Phe Thr Asn 195 200 205 Gly Leu Gly His Gly Val Asp LeuAsn His Ile Tyr Gly Glu Thr Leu 210 215 220 Ala Arg Gln Arg Lys Leu ArgLeu Phe Lys Asp Gly Lys Met Lys Tyr 225 230 235 240 Gln Ile Ile Asp GlyGlu Met Tyr Pro Pro Thr Val Lys Asp Thr Gln 245 250 255 Ala Glu Met IleTyr Pro Pro Gln Val Pro Glu His Leu Arg Phe Ala 260 265 270 Val Gly GlnGlu Val Phe Gly Leu Val Pro Gly Leu Met Met Tyr Ala 275 280 285 Thr IleTrp Leu Arg Glu His Asn Arg Val Cys Asp Val Leu Lys Gln 290 295 300 GluHis Pro Glu Trp Gly Asp Glu Gln Leu Phe Gln Thr Ser Arg Leu 305 310 315320 Ile Leu Ile Gly Glu Thr Ile Lys Ile Val Ile Glu Asp Tyr Val Gln 325330 335 His Leu Ser Gly Tyr His Phe Lys Leu Lys Phe Asp Pro Glu Leu Leu340 345 350 Phe Asn Lys Gln Phe Gln Tyr Gln Asn Arg Ile Ala Ala Glu PheAsn 355 360 365 Thr Leu Tyr His Trp His Pro Leu Leu Pro Asp Thr Phe GlnIle His 370 375 380 Asp Gln Lys Tyr Asn Tyr Gln Gln Phe Ile Tyr Asn AsnSer Ile Leu 385 390 395 400 Leu Glu His Gly Ile Thr Gln Phe Val Glu SerPhe Thr Arg Gln Ile 405 410 415 Ala Gly Arg Val Ala Gly Gly Arg Asn ValPro Pro Ala Val Gln Lys 420 425 430 Val Ser Gln Ala Ser Ile Asp Gln SerArg Gln Met Lys Tyr Gln Ser 435 440 445 Phe Asn Glu Tyr Arg Lys Arg PheMet Leu Lys Pro Tyr Glu Ser Phe 450 455 460 Glu Glu Leu Thr Gly Glu LysGlu Met Ser Ala Glu Leu Glu Ala Leu 465 470 475 480 Tyr Gly Asp Ile AspAla Val Glu Leu Tyr Pro Ala Leu Leu Val Glu 485 490 495 Lys Pro Arg ProAsp Ala Ile Phe Gly Glu Thr Met Val Glu Val Gly 500 505 510 Ala Pro PheSer Leu Lys Gly Leu Met Gly Asn Val Ile Cys Ser Pro 515 520 525 Ala TyrTrp Lys Pro Ser Thr Phe Gly Gly Glu Val Gly Phe Gln Ile 530 535 540 IleAsn Thr Ala Ser Ile Gln Ser Leu Ile Cys Asn Asn Val Lys Gly 545 550 555560 Cys Pro Phe Thr Ser Phe Ser Val Pro Asp Pro Glu Leu Ile Lys Thr 565570 575 Val Thr Ile Asn Ala Ser Ser Ser Arg Ser Gly Leu Asp Asp Ile Asn580 585 590 Pro Thr Val Leu Leu Lys Glu Arg Ser Thr Glu Leu 595 600 31474 PRT Homo sapiens 31 Met Ala Thr Asn Trp Gly Ser Leu Leu Gln Asp LysGln Gln Leu Glu 1 5 10 15 Glu Leu Ala Arg Gln Ala Val Asp Arg Ala LeuAla Glu Gly Val Leu 20 25 30 Leu Arg Thr Ser Gln Glu Pro Thr Ser Ser GluVal Val Ser Tyr Ala 35 40 45 Pro Phe Thr Leu Phe Pro Ser Leu Val Pro SerAla Leu Leu Glu Gln 50 55 60 Ala Tyr Ala Val Gln Met Asp Phe Asn Leu LeuVal Asp Ala Val Ser 65 70 75 80 Gln Asn Ala Ala Phe Leu Glu Gln Thr LeuSer Ser Thr Ile Lys Gln 85 90 95 Asp Asp Phe Thr Ala Arg Leu Phe Asp IleHis Lys Gln Val Leu Lys 100 105 110 Glu Gly Ile Ala Gln Thr Val Phe LeuGly Leu Asn Arg Ser Asp Tyr 115 120 125 Met Phe Gln Arg Ser Ala Asp GlySer Pro Ala Leu Lys Gln Ile Glu 130 135 140 Ile Asn Thr Ile Ser Ala SerPhe Gly Gly Leu Ala Ser Arg Thr Pro 145 150 155 160 Ala Val His Arg HisVal Leu Ser Val Leu Ser Lys Thr Lys Glu Ala 165 170 175 Gly Lys Ile LeuSer Asn Asn Pro Ser Lys Gly Leu Ala Leu Gly Ile 180 185 190 Ala Lys AlaTrp Glu Leu Tyr Gly Ser Pro Asn Ala Leu Val Leu Leu 195 200 205 Ile AlaGln Glu Lys Glu Arg Asn Ile Phe Asp Gln Arg Ala Ile Glu 210 215 220 AsnGlu Leu Leu Ala Arg Asn Ile His Val Ile Arg Arg Thr Phe Glu 225 230 235240 Asp Ile Ser Glu Lys Gly Ser Leu Asp Gln Asp Arg Arg Leu Phe Val 245250 255 Asp Gly Gln Glu Ile Ala Val Val Tyr Phe Arg Asp Gly Tyr Met Pro260 265 270 Arg Gln Tyr Ser Leu Gln Asn Trp Glu Ala Arg Leu Leu Leu GluArg 275 280 285 Ser His Ala Ala Lys Cys Pro Asp Ile Ala Thr Gln Leu AlaGly Thr 290 295 300 Lys Lys Val Gln Gln Glu Leu Ser Arg Pro Gly Met LeuGlu Met Leu 305 310 315 320 Leu Pro Gly Gln Pro Glu Ala Val Ala Arg LeuArg Ala Thr Phe Ala 325 330 335 Gly Leu Tyr Ser Leu Asp Val Gly Glu GluGly Asp Gln Ala Ile Ala 340 345 350 Glu Ala Leu Ala Ala Pro Ser Arg PheVal Leu Lys Pro Gln Arg Glu 355 360 365 Gly Gly Gly Asn Asn Leu Tyr GlyGlu Glu Met Val Gln Ala Leu Lys 370 375 380 Gln Leu Lys Asp Ser Glu GluArg Ala Ser Tyr Ile Leu Met Glu Lys 385 390 395 400 Ile Glu Pro Glu ProPhe Glu Asn Cys Leu Leu Arg Pro Gly Ser Pro 405 410 415 Ala Arg Val ValGln Cys Ile Ser Glu Leu Gly Ile Phe Gly Val Tyr 420 425 430 Val Arg GlnGlu Lys Thr Leu Val Met Asn Lys His Val Gly His Leu 435 440 445 Leu ArgThr Lys Ala Ile Glu His Ala Asp Gly Gly Val Ala Ala Gly 450 455 460 ValAla Val Leu Asp Asn Pro Tyr Pro Val 465 470 32 384 PRT Homo sapiens 32Met Lys Val Thr Ser Leu Asp Gly Arg Gln Leu Arg Lys Met Leu Arg 1 5 1015 Lys Glu Ala Ala Ala Arg Cys Val Val Leu Asp Cys Arg Pro Tyr Leu 20 2530 Ala Phe Ala Ala Ser Asn Val Arg Gly Ser Leu Asn Val Asn Leu Asn 35 4045 Ser Val Val Leu Arg Arg Ala Arg Gly Gly Ala Val Ser Ala Arg Tyr 50 5560 Val Leu Pro Asp Glu Ala Ala Arg Ala Arg Leu Leu Gln Glu Gly Gly 65 7075 80 Gly Gly Val Ala Ala Val Val Val Leu Asp Gln Gly Ser Arg His Trp 8590 95 Gln Lys Leu Arg Glu Glu Ser Ala Ala Arg Val Val Leu Thr Ser Leu100 105 110 Leu Ala Cys Leu Pro Ala Gly Pro Arg Val Tyr Phe Leu Lys GlyGly 115 120 125 Tyr Glu Thr Phe Tyr Ser Glu Tyr Pro Glu Cys Cys Val AspVal Lys 130 135 140 Pro Ile Ser Gln Glu Lys Ile Glu Ser Glu Arg Ala LeuIle Ser Gln 145 150 155 160 Cys Gly Lys Pro Val Val Asn Val Ser Tyr ArgPro Ala Tyr Asp Gln 165 170 175 Gly Gly Pro Val Glu Ile Leu Pro Phe LeuTyr Leu Gly Ser Ala Tyr 180 185 190 His Ala Ser Lys Cys Glu Phe Leu AlaAsn Leu His Ile Thr Ala Leu 195 200 205 Leu Asn Val Ser Arg Arg Thr SerGlu Ala Cys Met Thr His Leu His 210 215 220 Tyr Lys Trp Ile Pro Val GluAsp Ser His Thr Ala Asp Ile Ser Ser 225 230 235 240 His Phe Gln Glu AlaIle Asp Phe Ile Asp Cys Val Arg Glu Lys Gly 245 250 255 Gly Lys Val LeuVal His Cys Glu Ala Gly Ile Ser Arg Ser Pro Thr 260 265 270 Ile Cys MetAla Tyr Leu Met Lys Thr Lys Gln Phe Arg Leu Lys Glu 275 280 285 Ala PheAsp Tyr Ile Lys Gln Arg Arg Ser Met Val Ser Pro Asn Phe 290 295 300 GlyPhe Met Gly Gln Leu Leu Gln Tyr Glu Ser Glu Ile Leu Pro Ser 305 310 315320 Thr Pro Asn Pro Gln Pro Pro Ser Cys Gln Gly Glu Ala Ala Gly Ser 325330 335 Ser Leu Ile Gly His Leu Gln Thr Leu Ser Pro Asp Met Gln Gly Ala340 345 350 Tyr Cys Thr Phe Pro Ala Ser Val Leu Ala Pro Val Pro Thr HisSer 355 360 365 Thr Val Ser Glu Leu Ser Arg Ser Pro Val Ala Thr Ala ThrSer Cys 370 375 380 33 231 PRT Homo sapiens 33 Met Ala Gly Lys Lys ValLeu Ile Val Tyr Ala His Gln Glu Pro Lys 1 5 10 15 Ser Phe Asn Gly SerLeu Lys Asn Val Ala Val Asp Glu Leu Ser Arg 20 25 30 Gln Gly Cys Thr ValThr Val Ser Asp Leu Tyr Ala Met Asn Phe Glu 35 40 45 Pro Arg Ala Thr AspLys Asp Ile Thr Gly Thr Leu Ser Asn Pro Glu 50 55 60 Val Phe Asn Tyr GlyVal Glu Thr His Glu Ala Tyr Lys Gln Arg Ser 65 70 75 80 Leu Ala Ser AspIle Thr Asp Glu Gln Lys Lys Val Arg Glu Ala Asp 85 90 95 Leu Val Ile PheGln Phe Pro Leu Tyr Trp Phe Ser Val Pro Ala Ile 100 105 110 Leu Lys GlyTrp Met Asp Arg Val Leu Cys Gln Gly Phe Ala Phe Asp 115 120 125 Ile ProGly Phe Tyr Asp Ser Gly Leu Leu Gln Gly Lys Leu Ala Leu 130 135 140 LeuSer Val Thr Thr Gly Gly Thr Ala Glu Met Tyr Thr Lys Thr Gly 145 150 155160 Val Asn Gly Asp Ser Arg Tyr Phe Leu Trp Pro Leu Gln His Gly Thr 165170 175 Leu His Phe Cys Gly Phe Lys Val Leu Ala Pro Gln Ile Ser Phe Ala180 185 190 Pro Glu Ile Ala Ser Glu Glu Glu Arg Lys Gly Met Val Ala AlaTrp 195 200 205 Ser Gln Arg Leu Gln Thr Ile Trp Lys Glu Glu Pro Ile ProCys Thr 210 215 220 Ala His Trp His Phe Gly Gln 225 230 34 2201 PRT Homosapiens 34 Met Gly Ala Met Thr Gln Leu Leu Ala Gly Val Phe Leu Ala PheLeu 1 5 10 15 Ala Leu Ala Thr Glu Gly Gly Val Leu Lys Lys Val Ile ArgHis Lys 20 25 30 Arg Gln Ser Gly Val Asn Ala Thr Leu Pro Glu Glu Asn GlnPro Val 35 40 45 Val Phe Asn His Val Tyr Asn Ile Lys Leu Pro Val Gly SerGln Cys 50 55 60 Ser Val Asp Leu Glu Ser Ala Ser Gly Glu Lys Asp Leu AlaPro Pro 65 70 75 80 Ser Glu Pro Ser Glu Ser Phe Gln Glu His Thr Val AspGly Glu Asn 85 90 95 Gln Ile Val Phe Thr His Arg Ile Asn Ile Pro Arg ArgAla Cys Gly 100 105 110 Cys Ala Ala Ala Pro Asp Val Lys Glu Leu Leu SerArg Leu Glu Glu 115 120 125 Leu Glu Asn Leu Val Ser Ser Leu Arg Glu GlnCys Thr Ala Gly Ala 130 135 140 Gly Cys Cys Leu Gln Pro Ala Thr Gly ArgLeu Asp Thr Arg Pro Phe 145 150 155 160 Cys Ser Gly Arg Gly Asn Phe SerThr Glu Gly Cys Gly Cys Val Cys 165 170 175 Glu Pro Gly Trp Lys Gly ProAsn Cys Ser Glu Pro Glu Cys Pro Gly 180 185 190 Asn Cys His Leu Arg GlyArg Cys Ile Asp Gly Gln Cys Ile Cys Asp 195 200 205 Asp Gly Phe Thr GlyGlu Asp Cys Ser Gln Leu Ala Cys Pro Ser Asp 210 215 220 Cys Asn Asp GlnGly Lys Cys Val Asn Gly Val Cys Ile Cys Phe Glu 225 230 235 240 Gly TyrAla Gly Ala Asp Cys Ser Arg Glu Ile Cys Pro Val Pro Cys 245 250 255 SerGlu Glu His Gly Thr Cys Val Asp Gly Leu Cys Val Cys His Asp 260 265 270Gly Phe Ala Gly Asp Asp Cys Asn Lys Pro Leu Cys Leu Asn Asn Cys 275 280285 Tyr Asn Arg Gly Arg Cys Val Glu Asn Glu Cys Val Cys Asp Glu Gly 290295 300 Phe Thr Gly Glu Asp Cys Ser Glu Leu Ile Cys Pro Asn Asp Cys Phe305 310 315 320 Asp Arg Gly Arg Cys Ile Asn Gly Thr Cys Tyr Cys Glu GluGly Phe 325 330 335 Thr Gly Glu Asp Cys Gly Lys Pro Thr Cys Pro His AlaCys His Thr 340 345 350 Gln Gly Arg Cys Glu Glu Gly Gln Cys Val Cys AspGlu Gly Phe Ala 355 360 365 Gly Leu Asp Cys Ser Glu Lys Arg Cys Pro AlaAsp Cys His Asn Arg 370 375 380 Gly Arg Cys Val Asp Gly Arg Cys Glu CysAsp Asp Gly Phe Thr Gly 385 390 395 400 Ala Asp Cys Gly Glu Leu Lys CysPro Asn Gly Cys Ser Gly His Gly 405 410 415 Arg Cys Val Asn Gly Gln CysVal Cys Asp Glu Gly Tyr Thr Gly Glu 420 425 430 Asp Cys Ser Gln Leu ArgCys Pro Asn Asp Cys His Ser Arg Gly Arg 435 440 445 Cys Val Glu Gly LysCys Val Cys Glu Gln Gly Phe Lys Gly Tyr Asp 450 455 460 Cys Ser Asp MetSer Cys Pro Asn Asp Cys His Gln His Gly Arg Cys 465 470 475 480 Val AsnGly Met Cys Val Cys Asp Asp Gly Tyr Thr Gly Glu Asp Cys 485 490 495 ArgAsp Arg Gln Cys Pro Arg Asp Cys Ser Asn Arg Gly Leu Cys Val 500 505 510Asp Gly Gln Cys Val Cys Glu Asp Gly Phe Thr Gly Pro Asp Cys Ala 515 520525 Glu Leu Ser Cys Pro Asn Asp Cys His Gly Gln Gly Arg Cys Val Asn 530535 540 Gly Gln Cys Val Cys His Glu Gly Phe Met Gly Lys Asp Cys Lys Glu545 550 555 560 Gln Arg Cys Pro Ser Asp Cys His Gly Gln Gly Arg Cys ValAsp Gly 565 570 575 Gln Cys Ile Cys His Glu Gly Phe Thr Gly Leu Asp CysGly Gln His 580 585 590 Ser Cys Pro Ser Asp Cys Asn Asn Leu Gly Gln CysVal Ser Gly Arg 595 600 605 Cys Ile Cys Asn Glu Gly Tyr Ser Gly Glu AspCys Ser Glu Val Ser 610 615 620 Pro Pro Lys Asp Leu Val Val Thr Glu ValThr Glu Glu Thr Val Asn 625 630 635 640 Leu Ala Trp Asp Asn Glu Met ArgVal Thr Glu Tyr Leu Val Val Tyr 645 650 655 Thr Pro Thr His Glu Gly GlyLeu Glu Met Gln Phe Arg Val Pro Gly 660 665 670 Asp Gln Thr Ser Thr IleIle Gln Glu Leu Glu Pro Gly Val Glu Tyr 675 680 685 Phe Ile Arg Val PheAla Ile Leu Glu Asn Lys Lys Ser Ile Pro Val 690 695 700 Ser Ala Arg ValAla Thr Tyr Leu Pro Ala Pro Glu Gly Leu Lys Phe 705 710 715 720 Lys SerIle Lys Glu Thr Ser Val Glu Val Glu Trp Asp Pro Leu Asp 725 730 735 IleAla Phe Glu Thr Trp Glu Ile Ile Phe Arg Asn Met Asn Lys Glu 740 745 750Asp Glu Gly Glu Ile Thr Lys Ser Leu Arg Arg Pro Glu Thr Ser Tyr 755 760765 Arg Gln Thr Gly Leu Ala Pro Gly Gln Glu Tyr Glu Ile Ser Leu His 770775 780 Ile Val Lys Asn Asn Thr Arg Gly Pro Gly Leu Lys Arg Val Thr Thr785 790 795 800 Thr Arg Leu Asp Ala Pro Ser Gln Ile Glu Val Lys Asp ValThr Asp 805 810 815 Thr Thr Ala Leu Ile Thr Trp Phe Lys Pro Leu Ala GluIle Asp Gly 820 825 830 Ile Glu Leu Thr Tyr Gly Ile Lys Asp Val Pro GlyAsp Arg Thr Thr 835 840 845 Ile Asp Leu Thr Glu Asp Glu Asn Gln Tyr SerIle Gly Asn Leu Lys 850 855 860 Pro Asp Thr Glu Tyr Glu Val Ser Leu IleSer Arg Arg Gly Asp Met 865 870 875 880 Ser Ser Asn Pro Ala Lys Glu ThrPhe Thr Thr Gly Leu Asp Ala Pro 885 890 895 Arg Asn Leu Arg Arg Val SerGln Thr Asp Asn Ser Ile Thr Leu Glu 900 905 910 Trp Arg Asn Gly Lys AlaAla Ile Asp Ser Tyr Arg Ile Lys Tyr Ala 915 920 925 Pro Ile Ser Gly GlyAsp His Ala Glu Val Asp Val Pro Lys Ser Gln 930 935 940 Gln Ala Thr ThrLys Thr Thr Leu Thr Gly Leu Arg Pro Gly Thr Glu 945 950 955 960 Tyr GlyIle Gly Val Ser Ala Val Lys Glu Asp Lys Glu Ser Asn Pro 965 970 975 AlaThr Ile Asn Ala Ala Thr Glu Leu Asp Thr Pro Lys Asp Leu Gln 980 985 990Val Ser Glu Thr Ala Glu Thr Ser Leu Thr Leu Leu Trp Lys Thr Pro 995 10001005 Leu Ala Lys Phe Asp Arg Tyr Arg Leu Asn Tyr Ser Leu Pro Thr 10101015 1020 Gly Gln Trp Val Gly Val Gln Leu Pro Arg Asn Thr Thr Ser Tyr1025 1030 1035 Val Leu Arg Gly Leu Glu Pro Gly Gln Glu Tyr Asn Val LeuLeu 1040 1045 1050 Thr Ala Glu Lys Gly Arg His Lys Ser Lys Pro Ala ArgVal Lys 1055 1060 1065 Ala Ser Thr Glu Gln Ala Pro Glu Leu Glu Asn LeuThr Val Thr 1070 1075 1080 Glu Val Gly Trp Asp Gly Leu Arg Leu Asn TrpThr Ala Ala Asp 1085 1090 1095 Gln Ala Tyr Glu His Phe Ile Ile Gln ValGln Glu Ala Asn Lys 1100 1105 1110 Val Glu Ala Ala Arg Asn Leu Thr ValPro Gly Ser Leu Arg Ala 1115 1120 1125 Val Asp Ile Pro Gly Leu Lys AlaAla Thr Pro Tyr Thr Val Ser 1130 1135 1140 Ile Tyr Gly Val Ile Gln GlyTyr Arg Thr Pro Val Leu Ser Ala 1145 1150 1155 Glu Ala Ser Thr Gly GluThr Pro Asn Leu Gly Glu Val Val Val 1160 1165 1170 Ala Glu Val Gly TrpAsp Ala Leu Lys Leu Asn Trp Thr Ala Pro 1175 1180 1185 Glu Gly Ala TyrGlu Tyr Phe Phe Ile Gln Val Gln Glu Ala Asp 1190 1195 1200 Thr Val GluAla Ala Gln Asn Leu Thr Val Pro Gly Gly Leu Arg 1205 1210 1215 Ser ThrAsp Leu Pro Gly Leu Lys Ala Ala Thr His Tyr Thr Ile 1220 1225 1230 ThrIle Arg Gly Val Thr Gln Asp Phe Ser Thr Thr Pro Leu Ser 1235 1240 1245Val Glu Val Leu Thr Glu Glu Val Pro Asp Met Gly Asn Leu Thr 1250 12551260 Val Thr Glu Val Ser Trp Asp Ala Leu Arg Leu Asn Trp Thr Thr 12651270 1275 Pro Asp Gly Thr Tyr Asp Gln Phe Thr Ile Gln Val Gln Glu Ala1280 1285 1290 Asp Gln Val Glu Glu Ala His Asn Leu Thr Val Pro Gly SerLeu 1295 1300 1305 Arg Ser Met Glu Ile Pro Gly Leu Arg Ala Gly Thr ProTyr Thr 1310 1315 1320 Val Thr Leu His Gly Glu Val Arg Gly His Ser ThrArg Pro Leu 1325 1330 1335 Ala Val Glu Val Val Thr Glu Asp Leu Pro GlnLeu Gly Asp Leu 1340 1345 1350 Ala Val Ser Glu Val Gly Trp Asp Gly LeuArg Leu Asn Trp Thr 1355 1360 1365 Ala Ala Asp Asn Ala Tyr Glu His PheVal Ile Gln Val Gln Glu 1370 1375 1380 Val Asn Lys Val Glu Ala Ala GlnAsn Leu Thr Leu Pro Gly Ser 1385 1390 1395 Leu Arg Ala Val Asp Ile ProGly Leu Glu Ala Ala Thr Pro Tyr 1400 1405 1410 Arg Val Ser Ile Tyr GlyVal Ile Arg Gly Tyr Arg Thr Pro Val 1415 1420 1425 Leu Ser Ala Glu AlaSer Thr Ala Lys Glu Pro Glu Ile Gly Asn 1430 1435 1440 Leu Asn Val SerAsp Ile Thr Pro Glu Ser Phe Asn Leu Ser Trp 1445 1450 1455 Met Ala ThrAsp Gly Ile Phe Glu Thr Phe Thr Ile Glu Ile Ile 1460 1465 1470 Asp SerAsn Arg Leu Leu Glu Thr Val Glu Tyr Asn Ile Ser Gly 1475 1480 1485 AlaGlu Arg Thr Ala His Ile Ser Gly Leu Pro Pro Ser Thr Asp 1490 1495 1500Phe Ile Val Tyr Leu Ser Gly Leu Ala Pro Ser Ile Arg Thr Lys 1505 15101515 Thr Ile Ser Ala Thr Ala Thr Thr Glu Ala Leu Pro Leu Leu Glu 15201525 1530 Asn Leu Thr Ile Ser Asp Ile Asn Pro Tyr Gly Phe Thr Val Ser1535 1540 1545 Trp Met Ala Ser Glu Asn Ala Phe Asp Ser Phe Leu Val ThrVal 1550 1555 1560 Val Asp Ser Gly Lys Leu Leu Asp Pro Gln Glu Phe ThrLeu Ser 1565 1570 1575 Gly Thr Gln Arg Lys Leu Glu Leu Arg Gly Leu IleThr Gly Ile 1580 1585 1590 Gly Tyr Glu Val Met Val Ser Gly Phe Thr GlnGly His Gln Thr 1595 1600 1605 Lys Pro Leu Arg Ala Glu Ile Val Thr GluAla Glu Pro Glu Val 1610 1615 1620 Asp Asn Leu Leu Val Ser Asp Ala ThrPro Asp Gly Phe Arg Leu 1625 1630 1635 Ser Trp Thr Ala Asp Glu Gly ValPhe Asp Asn Phe Val Leu Lys 1640 1645 1650 Ile Arg Asp Thr Lys Lys GlnSer Glu Pro Leu Glu Ile Thr Leu 1655 1660 1665 Leu Ala Pro Glu Arg ThrArg Asp Leu Thr Gly Leu Arg Glu Ala 1670 1675 1680 Thr Glu Tyr Glu IleGlu Leu Tyr Gly Ile Ser Lys Gly Arg Arg 1685 1690 1695 Ser Gln Thr ValSer Ala Ile Ala Thr Thr Ala Met Gly Ser Pro 1700 1705 1710 Lys Glu ValIle Phe Ser Asp Ile Thr Glu Asn Ser Ala Thr Val 1715 1720 1725 Ser TrpArg Ala Pro Thr Ala Gln Val Glu Ser Phe Arg Ile Thr 1730 1735 1740 TyrVal Pro Ile Thr Gly Gly Thr Pro Ser Met Val Thr Val Asp 1745 1750 1755Gly Thr Lys Thr Gln Thr Arg Leu Val Lys Leu Ile Pro Gly Val 1760 17651770 Glu Tyr Leu Val Ser Ile Ile Ala Met Lys Gly Phe Glu Glu Ser 17751780 1785 Glu Pro Val Ser Gly Ser Phe Thr Thr Ala Leu Asp Gly Pro Ser1790 1795 1800 Gly Leu Val Thr Ala Asn Ile Thr Asp Ser Glu Ala Leu AlaArg 1805 1810 1815 Trp Gln Pro Ala Ile Ala Thr Val Asp Ser Tyr Val IleSer Tyr 1820 1825 1830 Thr Gly Glu Lys Val Pro Glu Ile Thr Arg Thr ValSer Gly Asn 1835 1840 1845 Thr Val Glu Tyr Ala Leu Thr Asp Leu Glu ProAla Thr Glu Tyr 1850 1855 1860 Thr Leu Arg Ile Phe Ala Glu Lys Gly ProGln Lys Ser Ser Thr 1865 1870 1875 Ile Thr Ala Lys Phe Thr Thr Asp LeuAsp Ser Pro Arg Asp Leu 1880 1885 1890 Thr Ala Thr Glu Val Gln Ser GluThr Ala Leu Leu Thr Trp Arg 1895 1900 1905 Pro Pro Arg Ala Ser Val ThrGly Tyr Leu Leu Val Tyr Glu Ser 1910 1915 1920 Val Asp Gly Thr Val LysGlu Val Ile Val Gly Pro Asp Thr Thr 1925 1930 1935 Ser Tyr Ser Leu AlaAsp Leu Ser Pro Ser Thr His Tyr Thr Ala 1940 1945 1950 Lys Ile Gln AlaLeu Asn Gly Pro Leu Arg Ser Asn Met Ile Gln 1955 1960 1965 Thr Ile PheThr Thr Ile Gly Leu Leu Tyr Pro Phe Pro Lys Asp 1970 1975 1980 Cys SerGln Ala Met Leu Asn Gly Asp Thr Thr Ser Gly Leu Tyr 1985 1990 1995 ThrIle Tyr Leu Asn Gly Asp Lys Ala Gln Ala Leu Glu Val Phe 2000 2005 2010Cys Asp Met Thr Ser Asp Gly Gly Gly Trp Ile Val Phe Leu Arg 2015 20202025 Arg Lys Asn Gly Arg Glu Asn Phe Tyr Gln Asn Trp Lys Ala Tyr 20302035 2040 Ala Ala Gly Phe Gly Asp Arg Arg Glu Glu Phe Trp Leu Gly Leu2045 2050 2055 Asp Asn Leu Asn Lys Ile Thr Ala Gln Gly Gln Tyr Glu LeuArg 2060 2065 2070 Val Asp Leu Arg Asp His Gly Glu Thr Ala Phe Ala ValTyr Asp 2075 2080 2085 Lys Phe Ser Val Gly Asp Ala Lys Thr Arg Tyr LysLeu Lys Val 2090 2095 2100 Glu Gly Tyr Ser Gly Thr Ala Gly Asp Ser MetAla Tyr His Asn 2105 2110 2115 Gly Arg Ser Phe Ser Thr Phe Asp Lys AspThr Asp Ser Ala Ile 2120 2125 2130 Thr Asn Cys Ala Leu Ser Tyr Lys GlyAla Phe Trp Tyr Arg Asn 2135 2140 2145 Cys His Arg Val Asn Leu Met GlyArg Tyr Gly Asp Asn Asn His 2150 2155 2160 Ser Gln Gly Val Asn Trp PheHis Trp Lys Gly His Glu His Ser 2165 2170 2175 Ile Gln Phe Ala Glu MetLys Leu Arg Pro Ser Asn Phe Arg Asn 2180 2185 2190 Leu Glu Gly Arg ArgLys Arg Ala 2195 2200 35 262 PRT Homo sapiens 35 Met Asp Pro Arg Leu SerThr Val Arg Gln Thr Cys Cys Cys Phe Asn 1 5 10 15 Val Arg Ile Ala ThrThr Ala Leu Ala Ile Tyr His Val Ile Met Ser 20 25 30 Val Leu Leu Phe IleGlu His Ser Val Glu Val Ala His Gly Lys Ala 35 40 45 Ser Cys Lys Leu SerGln Met Gly Tyr Leu Arg Ile Ala Asp Leu Ile 50 55 60 Ser Ser Phe Leu LeuIle Thr Met Leu Phe Ile Ile Ser Leu Ser Leu 65 70 75 80 Leu Ile Gly ValVal Lys Asn Arg Glu Lys Tyr Leu Leu Pro Phe Leu 85 90 95 Ser Leu Gln IleMet Asp Tyr Leu Leu Cys Leu Leu Thr Leu Leu Gly 100 105 110 Ser Tyr IleGlu Leu Pro Ala Tyr Leu Lys Leu Ala Ser Arg Ser Arg 115 120 125 Ala SerSer Ser Lys Phe Pro Leu Met Thr Leu Gln Leu Leu Asp Phe 130 135 140 CysLeu Ser Ile Leu Thr Leu Cys Ser Ser Tyr Met Glu Val Pro Thr 145 150 155160 Tyr Leu Asn Phe Lys Ser Met Asn His Met Asn Tyr Leu Pro Ser Gln 165170 175 Glu Asp Met Pro His Asn Gln Phe Ile Lys Met Met Ile Ile Phe Ser180 185 190 Ile Ala Phe Ile Thr Val Leu Ile Phe Lys Val Tyr Met Phe LysCys 195 200 205 Val Trp Arg Cys Tyr Arg Leu Ile Lys Cys Met Asn Ser ValGlu Glu 210 215 220 Lys Arg Asn Ser Lys Met Leu Gln Lys Val Val Leu ProSer Tyr Glu 225 230 235 240 Glu Ala Leu Ser Leu Pro Ser Lys Thr Pro GluGly Gly Pro Ala Pro 245 250 255 Pro Pro Tyr Ser Glu Val 260 36 729 PRTHomo sapiens 36 Met Gly Lys Lys Tyr Lys Asn Ile Val Leu Leu Lys Gly LeuGlu Val 1 5 10 15 Ile Asn Asp Tyr His Phe Arg Met Val Lys Ser Leu LeuSer Asn Asp 20 25 30 Leu Lys Leu Asn Leu Lys Met Arg Glu Glu Tyr Asp LysIle Gln Ile 35 40 45 Ala Asp Leu Met Glu Glu Lys Phe Arg Gly Asp Ala GlyLeu Gly Lys 50 55 60 Leu Ile Lys Ile Phe Glu Asp Ile Pro Thr Leu Glu AspLeu Ala Glu 65 70 75 80 Thr Leu Lys Lys Glu Lys Leu Lys Val Lys Gly ProAla Leu Ser Arg 85 90 95 Lys Arg Lys Lys Glu Val His Ala Thr Ser Pro AlaPro Ser Thr Ser 100 105 110 Ser Thr Val Lys Thr Glu Gly Ala Glu Ala ThrPro Gly Ala Gln Lys 115 120 125 Arg Lys Lys Ser Thr Lys Glu Lys Ala GlyPro Lys Gly Ser Lys Val 130 135 140 Ser Glu Glu Gln Thr Gln Pro Pro SerPro Ala Gly Ala Gly Met Ser 145 150 155 160 Thr Ala Met Gly Arg Ser ProSer Pro Lys Thr Ser Leu Ser Ala Pro 165 170 175 Pro Asn Ser Ser Ser ThrGlu Asn Pro Lys Thr Val Ala Lys Cys Gln 180 185 190 Val Thr Pro Arg ArgAsn Val Leu Gln Lys Arg Pro Val Ile Val Lys 195 200 205 Val Leu Ser ThrThr Lys Pro Phe Glu Tyr Glu Thr Pro Glu Met Glu 210 215 220 Lys Lys IleMet Phe His Ala Thr Val Ala Thr Gln Thr Gln Phe Phe 225 230 235 240 HisVal Lys Val Leu Asn Thr Ser Leu Lys Glu Lys Phe Asn Gly Lys 245 250 255Lys Ile Ile Ile Ile Ser Asp Tyr Leu Glu Tyr Asp Ser Leu Leu Glu 260 265270 Val Asn Glu Glu Ser Thr Val Ser Glu Ala Gly Pro Asn Gln Thr Phe 275280 285 Glu Val Pro Asn Lys Ile Ile Asn Arg Ala Lys Glu Thr Leu Lys Ile290 295 300 Asp Ile Leu His Lys Gln Ala Ser Gly Asn Ile Val Tyr Gly ValPhe 305 310 315 320 Met Leu His Lys Lys Thr Val Asn Gln Lys Thr Thr IleTyr Glu Ile 325 330 335 Gln Asp Asp Arg Gly Lys Met Asp Val Val Gly ThrGly Gln Cys His 340 345 350 Asn Ile Pro Cys Glu Glu Gly Asp Lys Leu GlnLeu Phe Cys Phe Arg 355 360 365 Leu Arg Lys Lys Asn Gln Met Ser Lys LeuIle Ser Glu Met His Ser 370 375 380 Phe Ile Gln Ile Lys Lys Lys Thr AsnPro Arg Asn Asn Asp Pro Lys 385 390 395 400 Ser Met Lys Leu Pro Gln GluGln Arg Gln Leu Pro Tyr Pro Ser Glu 405 410 415 Ala Ser Thr Thr Phe ProGlu Ser His Leu Arg Thr Pro Gln Met Pro 420 425 430 Pro Thr Thr Pro SerSer Ser Phe Phe Thr Lys Lys Ser Glu Asp Thr 435 440 445 Ile Ser Lys MetAsn Asp Phe Met Arg Met Gln Ile Leu Lys Glu Gly 450 455 460 Ser His PhePro Gly Pro Phe Met Thr Ser Ile Gly Pro Ala Glu Ser 465 470 475 480 HisPro His Thr Pro Gln Met Pro Pro Ser Thr Pro Ser Ser Ser Phe 485 490 495Leu Thr Thr Leu Lys Pro Arg Leu Lys Thr Glu Pro Glu Glu Val Ser 500 505510 Ile Glu Asp Ser Ala Gln Ser Asp Leu Lys Glu Val Met Val Leu Asn 515520 525 Ala Thr Glu Ser Phe Val Tyr Glu Pro Lys Glu Gln Lys Lys Met Phe530 535 540 His Ala Thr Val Ala Thr Glu Asn Glu Val Phe Arg Val Lys ValPhe 545 550 555 560 Asn Ile Asp Leu Lys Glu Lys Phe Thr Pro Lys Lys IleIle Ala Ile 565 570 575 Ala Asn Tyr Val Cys Arg Asn Gly Phe Leu Glu ValTyr Pro Phe Thr 580 585 590 Leu Val Ala Asp Val Asn Ala Asp Arg Asn MetGlu Ile Pro Lys Gly 595 600 605 Leu Ile Arg Ser Ala Ser Val Thr Pro LysIle Asn Gln Leu Cys Ser 610 615 620 Gln Thr Lys Gly Ser Phe Val Asn GlyVal Phe Glu Val His Lys Lys 625 630 635 640 Asn Val Arg Gly Glu Phe ThrTyr Tyr Glu Ile Gln Asp Asn Thr Gly 645 650 655 Lys Met Glu Val Val ValHis Gly Arg Leu Asn Thr Ile Asn Cys Glu 660 665 670 Glu Gly Asp Lys LeuLys Leu Thr Ser Phe Glu Leu Ala Pro Lys Ser 675 680 685 Gly Asn Thr GlyGlu Leu Arg Ser Val Ile His Ser His Ile Lys Val 690 695 700 Ile Lys ThrArg Lys Asn Lys Lys Asp Ile Leu Asn Pro Asp Ser Ser 705 710 715 720 MetGlu Thr Ser Pro Asp Phe Phe Phe 725 37 354 PRT Homo sapiens 37 Met ArgLeu Ala Val Leu Phe Ser Gly Ala Leu Leu Gly Leu Leu Ala 1 5 10 15 AlaGln Gly Thr Gly Asn Asp Cys Pro His Lys Lys Ser Ala Thr Leu 20 25 30 LeuPro Ser Phe Thr Val Thr Pro Thr Val Thr Glu Ser Thr Gly Thr 35 40 45 ThrSer His Arg Thr Thr Lys Ser His Lys Thr Thr Thr His Arg Thr 50 55 60 ThrThr Thr Gly Thr Thr Ser His Gly Pro Thr Thr Ala Thr His Asn 65 70 75 80Pro Thr Thr Thr Ser His Gly Asn Val Thr Val His Pro Thr Ser Asn 85 90 95Ser Thr Ala Thr Ser Gln Gly Pro Ser Thr Ala Thr His Ser Pro Ala 100 105110 Thr Thr Ser His Gly Asn Ala Thr Val His Pro Thr Ser Asn Ser Thr 115120 125 Ala Thr Ser Pro Gly Phe Thr Ser Ser Ala His Pro Glu Pro Pro Pro130 135 140 Pro Ser Pro Ser Pro Ser Pro Thr Ser Lys Glu Thr Ile Gly AspTyr 145 150 155 160 Thr Trp Thr Asn Gly Ser Gln Pro Cys Val His Leu GlnAla Gln Ile 165 170 175 Gln Ile Arg Val Met Tyr Thr Thr Gln Gly Gly GlyGlu Ala Trp Gly 180 185 190 Ile Ser Val Leu Asn Pro Asn Lys Thr Lys ValGln Gly Ser Cys Glu 195 200 205 Gly Ala His Pro His Leu Leu Leu Ser PhePro Tyr Gly His Leu Ser 210 215 220 Phe Gly Phe Met Gln Asp Leu Gln GlnLys Val Val Tyr Leu Ser Tyr 225 230 235 240 Met Ala Val Glu Tyr Asn ValSer Phe Pro His Ala Ala Lys Trp Thr 245 250 255 Phe Ser Ala Gln Asn AlaSer Leu Arg Asp Leu Gln Ala Pro Leu Gly 260 265 270 Gln Ser Phe Ser CysSer Asn Ser Ser Ile Ile Leu Ser Pro Ala Val 275 280 285 His Leu Asp LeuLeu Ser Leu Arg Leu Gln Ala Ala Gln Leu Pro His 290 295 300 Thr Gly ValPhe Gly Gln Ser Phe Ser Cys Pro Ser Asp Arg Ser Ile 305 310 315 320 LeuLeu Pro Leu Ile Ile Gly Leu Ile Leu Leu Gly Leu Leu Ala Leu 325 330 335Val Leu Ile Ala Phe Cys Ile Ile Arg Arg Arg Pro Ser Ala Tyr Gln 340 345350 Ala Leu 38 351 PRT Homo sapiens 38 Met Pro Gly Ser Ala Ala Lys GlySer Glu Leu Ser Glu Arg Ile Glu 1 5 10 15 Ser Phe Val Glu Thr Leu LysArg Gly Gly Gly Pro Arg Ser Ser Glu 20 25 30 Glu Met Ala Arg Glu Thr LeuGly Leu Leu Arg Gln Ile Ile Thr Asp 35 40 45 His Arg Trp Ser Asn Ala GlyGlu Leu Met Glu Leu Ile Arg Arg Glu 50 55 60 Gly Arg Arg Met Thr Ala AlaGln Pro Ser Glu Thr Thr Val Gly Asn 65 70 75 80 Met Val Arg Arg Val LeuLys Ile Ile Arg Glu Glu Tyr Gly Arg Leu 85 90 95 His Gly Arg Ser Asp GluSer Asp Gln Gln Glu Ser Leu His Lys Leu 100 105 110 Leu Thr Ser Gly GlyLeu Asn Glu Asp Phe Ser Phe His Tyr Ala Gln 115 120 125 Leu Gln Ser AsnIle Ile Glu Ala Ile Asn Glu Leu Leu Val Glu Leu 130 135 140 Glu Gly ThrMet Glu Asn Ile Ala Ala Gln Ala Leu Glu His Ile His 145 150 155 160 SerAsn Glu Val Ile Met Thr Ile Gly Phe Ser Arg Thr Val Glu Ala 165 170 175Phe Leu Lys Glu Ala Ala Arg Lys Arg Lys Phe His Val Ile Val Ala 180 185190 Glu Cys Ala Pro Phe Cys Gln Gly His Glu Met Ala Val Asn Leu Ser 195200 205 Lys Ala Gly Ile Glu Thr Thr Val Met Thr Asp Ala Ala Ile Phe Ala210 215 220 Val Met Ser Arg Val Asn Lys Val Ile Ile Gly Thr Lys Thr IleLeu 225 230 235 240 Ala Asn Gly Ala Leu Arg Ala Val Thr Gly Thr His ThrLeu Ala Leu 245 250 255 Ala Ala Lys His His Ser Thr Pro Leu Ile Val CysAla Pro Met Phe 260 265 270 Lys Leu Ser Pro Gln Phe Pro Asn Glu Glu AspSer Phe His Lys Phe 275 280 285 Val Ala Pro Glu Glu Val Leu Pro Phe ThrGlu Gly Asp Ile Leu Glu 290 295 300 Lys Val Ser Val His Cys Pro Val PheAsp Tyr Val Pro Pro Glu Leu 305 310 315 320 Ile Thr Leu Phe Ile Ser AsnIle Gly Gly Asn Ala Pro Ser Tyr Ile 325 330 335 Tyr Arg Leu Met Ser GluLeu Tyr His Pro Asp Asp His Val Leu 340 345 350

We claim:
 1. A method for detecting breast cancer in a subject, saidmethod comprising the steps of: (a) contacting a biological sample fromthe subject with an agent that binds to a polypeptide comprising anamino acid sequence recited in any one of SEQ ID NOS:20-38; (b)determining a level of binding of said agent to said polypeptide; (c)comparing the level of binding of said agent to said polypeptide to acontrol level of binding; and (d) producing a diagnosis based on aresult from step (c).
 2. The method of claim 1, wherein said agent is anantibody directed against said polypeptide.
 3. The method of claim 2,wherein the antibody is selected from the group consisting of Fabfragment, Fab₂ fragment, single chain antibody, chimeric antibody,monoclonal antibody and polyclonal antibody.
 4. The method of claim 1,wherein the level of binding of said agent to said polypeptide in saidbiological sample is determined using a technology selected from thegroup consisting of ELISA, microarray technology, and biochiptechnology.
 5. The method of claim 1, wherein said agent binds to apolypeptide comprising an amino acid sequence recited in SEQ ID NO:29.6. A method for detecting breast cancer in a subject, said methodcomprising the steps of: (a) determining a level of a transcribedpolynucleotide in a biological sample from said subject, wherein saidtranscribed polynucleotide comprises a nucleic acid sequence recited inany one of SEQ ID NOS:1-19, or a complement of any of the foregoingnucleic acid sequences; (b) comparing the level of said transcribedpolynucleotide in said biological sample to a control level of saidtranscribed polynucleotide; and (c) producing a diagnosis based on aresult from step (b).
 7. The method of claim 6, wherein said transcribedpolynucleotide is an mRNA, and wherein the level of mRNA in saidbiological sample is determined using a method selected from the groupconsisting of Northern hybridization, RT-PCR, microarray technology, andbiochip technology.
 8. The method of claim 6, wherein the transcribedpolynucleotide comprises a nucleic acid sequence recited in SEQ IDNO:10, or a complement thereof.
 9. A method for detecting breast cancerin a subject, said method comprising the steps of: (a) determining anexpression pattern of two or more breast cancer-specific markers in abiological sample from said subject; (b) comparing the expressionpattern of the two or more breast cancer-specific markers in saidbiological sample to a control expression pattern; and (c) producing adiagnosis based on a result from step (b), wherein said breastcancer-specific marker is a polynucleotide comprising a nucleic acidsequence recited in any one of SEQ ID NOS:1-19 or a polypeptidecomprising an amino acid sequence recited in any one of SEQ IDNOS:20-38.
 10. The method of claim 9, wherein the expression pattern oftranscribed polynucleotides in the biological sample is determined usinga method selected from the group consisting of Northern hybridizationand RT-PCR.
 11. The method of claim 9, wherein the expression pattern ofpolypeptides in the biological sample is determined using antibodiesdirected against the polypeptides.
 12. The method of claim 9, whereinthe expression pattern of two or more breast cancer-specific markers isdetermined using microarray or biochip technology.
 13. A pharmaceuticalcomposition for preventing or treating breast cancer, comprisingpharmaceutically acceptable carrier and an agent capable of modulatingan activity of a breast cancer-specific marker or an expression level ofa breast cancer-specific gene, wherein said breast cancer-specificmarker is a polynucleotide comprising a nucleic acid sequence recited inany one of SEQ ID NOS:1-19 or a polypeptide comprising an amino acidsequence recited in any one of SEQ ID NOS:20-38, and wherein said breastcancer-specific gene is any one of the genes listed in Tables 4 and 5.14. A method for preventing or treating breast cancer in a subject, saidmethod comprising the step of: introducing into the subject an effectiveamount of the pharmaceutical composition of claim
 13. 15. A method ofidentifying an agent capable of binding to a breast cancer-specificmarker, said method comprising: contacting a breast cancer-specificmarker with a candidate agent; and determining a binding affinity ofsaid candidate agent to said breast cancer-specific marker, wherein saidbreast cancer-specific marker is a polynucleotide comprising a nucleicacid sequence recited in any one of SEQ ID NOS:1-19 or a polypeptidecomprising an amino acid sequence recited in any one of SEQ IDNOS:20-38.
 16. The method of claim 15, wherein the breastcancer-specific marker or the candidate agent contains a label.
 17. Amethod of identifying an agent capable of modulating an activity of abreast cancer-specific marker, comprising: contacting a breastcancer-specific marker with a candidate agent; determining an activityof said breast cancer-specific marker in the presence of said candidateagent; determining the activity of said breast cancer-specific marker inthe absence of said candidate agent; and determining whether saidcandidate agent affects the activity of said breast cancer-specificmarker, wherein said breast cancer-specific marker is a polynucleotidecomprising a nucleic acid sequence recited in any one of SEQ ID NOS:1-19or a polypeptide comprising an amino acid sequence recited in any one ofSEQ ID NOS:20-38.
 18. A biochip comprising any one of: (a) apolynucleotide comprising a nucleic acid sequence recited in any one ofSEQ ID NOS:1-19; (b) a variant of the polynucleotides of (a); (c) apolypeptide comprising an amino acid sequence recited in any one of SEQID NOS:20-38; and (d) a variant of the polypeptide of (c), wherein thebiochip is utilized for diagnosing breast cancer or screening agentsthat inhibit breast cancer.
 19. A kit for diagnosing breast cancer, saidkit comprising a polynucleotide probe or an antibody, wherein saidpolynucleotides probe specifically binds to a transcribed polynucleotidecomprising a nucleic acid sequence recited in any one of SEQ IDNOS:1-19, and wherein said antibody is capable of immunospecific bindingto a polypeptide comprising an amino acid sequence recited in any one ofSEQ ID NOS:20-38.
 20. The kit of claim 19, wherein the polynucleotidesprobe specifically binds to a transcribed polynucleotide comprising anucleic acid sequence recited in SEQ ID NO:10, and wherein the antibodyis capable of immunospecific binding to a polypeptide comprising anamino acid sequence recited in SEQ ID NO:29.